Oral Med Pathol 12 (2008) 57

Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology

Date: November 17-18, 2007 Venue: Howard International House, Taipei, the Republic of China

Oraganizaing Committee: President: Chun-Pin Chiang, School of dentistry, College of Medicine, National Taiwan University Vice Presidents: Li-Min Lin, College of Dental Medicine kaohsiung Medical University, Ying-Tai Jin, National Cheng Kung University Secretary General: Ying-Tai Jin, Nationatl Cheng Kung University Organizers: Taiwan Academy of Oral Pathology and School of Dentistry of Medicine, National Taiwan University

myoepithelial and/or basal cells, glycogen-rich clear cells, Special lecture squamous epithelial cells and oncocytic cells may also be found focally or extensively in different tumour types. 1. Classification and diagnosis of salivary gland Grading of malignancy: The presence of many low-grade tumors based on the revised WHO classification and intermediate-grade carcinomas, which show greater Hiromasa Nikai propensity for the indolent aggression, and the ability of Hiroshima University, Hiroshima, Japan long standing benign tumours to undergo malignant transformation are striking features of salivary carcinomas. (1) General remarks of salivary gland tumours Therefore, the most important work for pathologists at We oral pathologists often have difficulties and confuse diagnosis of salivary gland neoplasms is to distinguish these in diagnosis of salivary gland tumours. This will be low- or intermediate grade carcinomas from both benign and attributable to our lack of accumulated experience due to highly aggressive tumour types often bearing microscopic their overall rarity, comprising only about 1% of human resemblances for determining the choice of treatments. tumours and less than 5% of all head and neck tumours. Their extreme morphological diversity between different (2) Classification of salivary gland tumours tumour types also makes diagnosis difficult. Despite the The same classification is applied to major and minor rather small number of cases encountered in routine salivary gland tumours. The WHO classification (1991) histologic examination, there are so many tumour entities. revealed a great expansion in the histologic types recognized Histologic patterns and cell components are often shared compared with the 1976 classification. In addition to among a variety of salivary adenomas and carcinomas with the recognition of many new types during the past two different clinical behaviors and prognosis. decades, as the most attributable reason for this significant Anatomical sites: Some tumours occur exclusively or increase of named entities, a variety of carcinomas, which predominantly in the parotid gland, whereas some are had been lumped together under the common heading of essentially limited to the minor salivary glands. This “”, had been clarified to be specific entities knowledge may guide diagnosis. with distinct histology and biologic behaviors and these Histological features and cell components: Reflecting a were classified individually. normal histoarchitecture of the tissue of origin, many of the In the newly published WHO classification (2005), there salivary gland tumours show the biphasic structure composed are not so many significant changes when compared with the of inner luminal epithelial cells and outer non-luminal 1991 classification. The revised classification is almost same myoepithelial or basaloid cells. On the other hand, some to the classification described in the AFIP atlas (1995). tumours may be almost purely epithelial or myoepithelial in (1) Tumours are divided into two groups, malignant cellular composition. The histological diversity is often epithelial tumours and benign epithelial tumours. brought by the neoplastic myoepithelial cells, as an important (2) Clear cell carcinoma, not otherwise specified, low- morphologic modifier, which exhibit a wide spectrum of grade cribriform cystadenocarcinoma and sialo- phenotypic expression. These “modified” myoepithelial or blastoma are newly adopted in the malignant epithe- basaloid cells often produce glycosaminoglycans and basal lial tumour group. lamina materials that may form the cribriform pseudoluminae. (3) As minor changes in the malignant epithelial tumour Cribriform pattern may also be formed by true ductal spaces. group, adenocarcinoma is defined as adenocarci- Biphasic tubular and cribriform structures are shared by a , not otherwise specified. Cystadenocarci- variety of tumour types. In addition to the cell components noma includes not only papillary cystadenocarcinoma showing differentiation toward the acinar, ductal, but also non-papillary cystadenocarcinoma. Carci- 58 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology

noma ex , carcinosarcoma carcinoma, , small cell carcinoma and and metastasizing pleomorphic adenoma are indi- some non-sialogenic tumours. Polymorphous low- vidually classified and their comprehensive heading grade adenocarcinoma is a distinctive carcinoma of “carcinoma in pleomorphic adenoma” (malignant with low metastatic potential nearly always mixed tumour) was excluded. Large cell carcinoma involving minor salivary glands, whereas salivary and lymphoepithelial carcinoma are also indepen- duct carcinoma is one of the most aggressive dently classified without their common heading of adeno­carcinomas mainly involving the parotid. In “undifferentiated carcinoma” previously used. recent years, the presence of low-grade variant of (4) In the benign epithelial tumour group, sebaceous this carcinoma has been suggested. Low-grade and non-sebaceous lymphadenomas are classified cribriform cystadenocarcinoma may be regarded to under the heading of lymphadenoma. Cystadenoma represent low-grade salivary duct carcinoma. includes papillary cystadenoma and mucinous cysta- denoma both of which were previously listed up as 2. Human papillomavirus in oral lesions and DNA subclassification. vaccines (5) Various synonyms previously used, such as myoepi- Hatsuhiko Maeda thelial adenoma, adenolymphoma, oncocytic adenoma, Department of Pathology, School of Dentistry, Aichi-Gakuin terminal duct adenocarcinoma and malignant myo- University, Nagoya, Japan epithelioma are not used in the present classification. In the past 20 years, there has been an increasing interest in human papillomaviruses (HPV) because of (3) Differential diagnosis of salivary gland tumours their potential role in the pathogenesis of malignant Discussion will be focused in the differential diagnosis tumors and HPV infections have been demonstrated in of (1) tumours mainly composed of clear cells, (2) tumours several oral disorders. In a recent meta-analysis, HPV mainly composed of oncocytic cells, (3) tumours with basal/ was indeed confirmed as an independent risk factor for basaloid cells as essential component, and (4) tumours oral carcinoma. To date, totally more than 100 types of showing cribriform pattern in their histology. Diagnostic HPV have been identified. As in anogenital cancers, HPV approach to these tumour groups will be discussed. Concept type 16 is the most prevalent type in oral carcinomas. The and diagnostic criteria of the newly classified tumour benign oral lesions, associated with HPV infection, include types will be explained here. The presentation will also , papilloma, condyloma acuminatum, verruca highlight an update on the conceptual, histopathologic vulgaris, and focal epithelial hyperplasia (FEH). Papillomas and clinicopathologic aspects and, if possible, immuno­ and condylomas are mostly caused by HPV type 6 or 11, histochemical and ultrastructural features of each tumour while oral verrucas are associated with the skin types 2 or entity included in these groups. 4. The FEH lesions are caused by HPV types 13 and 32, (1) Salivary clear cell tumours include clear cell only detected in oral epithelium. In immunocompromised , sebaceous adenoma/carcinoma, clear patients, benign HPV-induced lesions are characterized cell variants of myoepithelioma/myoepithelial carci­ by atypical morphology and the simultaneous detection of noma, and acinic cell multiple HPV types. Oral benign HPV lesions are mostly carcinoma, monomorphic variant of epithelial- asymptomatic, and may persist or regress spontaneously. myoepithelial carcinoma, and clear cell carcinoma, In a previous investigation, we developed a novel NOS (hyalinizing clear cell carcinoma). hamster oral papillomavirus (HOPV) associated carcino­ (2) Oncocytic variants are also known to be present in genesis model by combining DMBA application with mucoepidermoid carcinoma, physical wounding of the hamster lingual mucosa. Using and myoepithelioma/myoepithelial carcinoma. Fea­ this animal model, we demonstrated that immunization with tures of other oncocytic tumours such as oncocytoma/ L1, E6 and E7 DNA vaccines prevent development of oncocytic carcinoma, Warthin tumour and oncocytic papillomavirus-associated oral carcinoma. Recently it has cystadenoma/cystadenocarcinoma are also briefly become clear that more potent methods for DNA vaccine outlined. delivery need to be developed to enhance the efficacy of (3) Included in the basaloid cell tumour group are basal DNA vaccines. Using this HOPV hamster model to compare cell adenoma/adenocarcinoma, adenoid cystic carci­ the anti-tumor effectiveness of different procedures of DNA noma and basaloid squamous cell carcinoma. vaccine delivery, intramuscular injection alone or with Sialoblastoma is one of the basaloid cell tumours as electroporation, we explained that in vivo electroporation is indicated by its synonyms such as congenital basal a potent method for DNA vaccine delivery. cell adenoma and basaloid adenocarcinoma. (4) Cribriform structure resembling that in is frequently or less frequently encountered in pleomorphic adenoma, / adenocarcinoma, polymorphous low-grade adeno­ carcinoma, myoepithelioma/myoepithelial carcinoma, salivary duct carcinoma, basaloid squamous cell Oral Med Pathol 12 (2008) 59

3. Parenchymal-stromal switching for extracellular positive and negative cell cycle regulators. Here we present matrix production: conceptual evidence for the our recent studies on abnormal expression of p27 and Aurora histopathological judgment of micro-invasion of A in oral squamous cell carcinoma (OSCC) that was oral squamous cell carcinomas achieved to examine their roles in OSCC. Takashi Saku p27 is a G1/S cyclin-dependent kinase inhibitor. Increase Division of Oral Pathology, Niigata University, Graduate in the cellular abundance of p27 upon induction of cell School of Medical and Dental Sciences, Niigata, Japan quiescence is primarily due to a decrease in the rate of its One of the most difficult issues in histopathological degradation. p27 is poly-ubiquitinated both in vivo and in diagnoses of squamous cell carcinoma (SCC) with superficial vitro and a lower amount of p27 ubiquitinating activity is spread or carcinoma in situ of the is the present in proliferating cells compared to quiescent cells. judgment between micro-invasion and crosscutting. To Furthermore, p27 ubiquitination requires its phosphorylation dissolve this issue, we have investigated the production of on threonine 187. Reduced expression of p27 was frequently extracellular matrix (ECM) molecules by carcinoma cells found in various cancers, and the lack of p27 is suggested to and stromal fibroblasts in different culture conditions as well be due to an enhancement of its degradation. We found that as in tissue sections. Immunohistochemically or hybrid- reduced expression of p27 was observed in about 90% of histochemically, we have been successful in demonstration OSCC cases examined and was well correlated with its of ECM deposition and production of epithelial cells in malignancy. Reduced p27 levels can be a powerful prognostic epithelial dysplasia or carcinoma in situ, which reflected in marker for poor survival in cancer patients. “loss of intercellular adherence,” one of the characteristic Recently, it has been reported that Skp2, an F-box histological features of epithelial dysplasia (WHO). How­ protein necessary for DNA replication, is required for the ever, once carcinoma cells start to invade, they stop to ubiquitination and consequent degradation of p27. A protein produce ECM by themselves, while the ECM production is called Cks1 is required for the Skp2-mediated ubiquitination taken over by stromal fibroblasts. The phenomena have been of p27. We, therefore, examined the correlation between confirmed by co-culture experiments of SCC cells and Skp2, Cks1 and p27. High expression of Skp2 was present in fibroblasts for their expression levels for ECM with PCR, 49 % of OSCC cases and significantly correlated with poor immunofluorescence, or immunoprecipitation, in which prognosis of OSCC patients. We also found inverse fibroblasts produced more ECM molecules, such as perlecan, correlation between the expression of Skp2 and p27 in tenascin, fibronectin, and laminin, and SCC cells less when OSCC. Interestingly, OSCC tissues with Skp2 expression compared with their singular cultivations. We call this had high p27 degradation activity. High expression of Cks1 phenomenon “switching of stromal induction from paren­ was present in 62% of OSCC. Cks1 expression was well chymal cells to stromal cells.” When applied this idea to correlated with Skp2 expression and poor prognosis, but not tissue sections of superficial carcinoma cases by immuno­ p27 expression. Cks1 overexpression enhanced Skp2 depen­ histochemistry for ECM, micro-invasive foci can be revealed dent p27 degradation. Silencing of either Skp2 or Cks1 by by the presence of ECM around the foci, while cross-cuts of siRNA oligos induced p27 accumulation and inhibited the epithelial projections are not surrounded by newly growth of OSCC cells both in vitro and in vivo. synthesized neoplastic stromal spaces, which is now an The serine/threonine kinase Aurora-A (Aur-A) is important aid for the determination of micro-invasion. essential for mitotic entry, centrosome duplication, spindle Based on these lines of evidence, we have proposed a formation, chromosome segregation and cytokinesis. Aur-A concept of “intra-epithelial stroma.” The intra-epithelial is a proto-oncoprotein overexpressed in a wide range of stroma is enhanced in epithelial dysplasia or carcinoma in human cancers. Overexpression of Aur-A is thought to be situ or in the characteristic stellate reticulum-like foci of caused by gene amplification or mRNA overexpression. ameloblastomas. We consider that ECM molecules are However, recent evidence revealed that the discrepancies required in their proliferation by tumor cells. This is also the between amplification of Aur-A and overexpression rates of case in the process of metastasis, because carcinoma cell Aur-A mRNA were observed in various cancers. We also systems with highly metastatic potentials including ACCM found OSCC exhibited overexpression and stabilization of have been shown to express higher levels of ECM molecule Aur-A protein without gene amplification or mRNA genes when analyzed by DNA micro-array as well as real overexpression. Therefore, we tested the hypothesis that time PCR. aberration of the protein destruction system induces accumulation and consequently overexpression of Aur-A in 4. Abnormal degradation of cell cycle regulators, cancer. Aur-A protein was ubiquitinylated by APCCdh1 and p27 and Aurora A, in consequently degraded when cells exited mitosis, and Takashi Takata, Yasusei Kudo, Shojiro Kitajima, Ikuko phosphorylation of Aur-A on Ser51 was observed during Ogawa, Mutsumi Miyauchi mitosis. Phosphorylation of Aur-A on Ser51 inhibited its Department of Oral and Maxillofacial Pathobiology, Graduate APCCdh1-mediated ubiquitylation and consequent degrad­ School of Biomedical Sciences, Hiroshima University, Hiro­ ation. Interestingly, constitutive phosphorylation on Ser51 shima, Japan was observed in OSCC cells with protein overexpression Proliferation and progression of cancer cells have been and stabilization. Indeed, phosphorylation on Ser51 was known to closely relate to the abnormalities of various observed in OSCC tissues with Aur-A protein overexpression. 60 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology

Moreover, an Aur-A Ser51 phospho-mimetic mutant displayed Twist expression and induce mesenchymal-epithelial stabilization of protein during cell cycle pro­gression and transition (MET) in oral cancer cells. Inhibition of enhanced ability to cell transformation. These findings phosphorylated Akt by PIA effectively increased GSK3 suggest that p27 down-regulation may be caused by Skp2 phosphorylation and decreased NFkB and JNK signaling, and Cks1 overexpression in OSCC and there is a new mode whereas it did not affect phosphorylation of ERK. Akt of Aur-A overexpression in cancer through phos­phorylation- inhibition led to downregulation of Snail and Twist dependent inhibition of its proteolysis in addition to gene expression, induced E-cadherin expression, and reduced amplification and mRNA overexpression. Overall, we tumor cell migration. Interestingly, Akt inhibition did not believe that inhibition of p27 degradation and Aur-A affect SIP-1/ZEB-2 expression. All of these findings suggest phosphorylation can be a novel target for OSCC therapy. that Akt inhibition could induce the MET through downregulation of Snail and Twist, not SIP-1/ZEB-2 in 5. The role of epithelial-mesenchymal transition OSCC cells, and the strategy of Akt inhibition might be one (EMT) in oral cancer of useful therapeutic tools in the treatment of oral cancer Seong-Doo Hong patients. Department of Oral Pathology, School of Dentistry, Seoul National University, Seoul, Korea 6. Potential roles of heme oxygenase-1 in , Oral squamous cell carcinoma (OSCC) is the most periodontitis, oral mucosal disease and oral cancer common neoplasm of the head and neck. Carcinoma cells Eun-Cheol Kim accumulate a series of genetic and/or epigenetic changes and Department of Oral & Maxillofacial Pathology, Wonkwang altered phenotypes during tumor progression. Loss of the University, Iksan, South Korea epithelial morphology and acquisition of mesenchymal Heme oxygenase (HO) is the rate-limiting enzyme in characteristics, epithelial-mesenchymal transition (EMT), is heme catabolism and converts heme to biliverdin, carbon typical for carcinoma cells during tumor progression and monoxide (CO), and free iron. One of three mammalian HO correlates with local invasiveness and metastatic potential of isoforms, HO-1 (also called heat shock protein 32), is a the tumor. Such changes are associated with aberrant stress-responsive protein induced by various agents and is expression of developmentally regulated and mesenchymal involved in a variety of regulatory and protective mechanisms cell-related genes. Among the mechanisms largely associated in cells. Increases in HO-1 have been detected in inflammatory with the metastatic conversion of epithelial cells and the diseases such as allergic asthma and rhinitis. However, EMT, one of the prominent changes is the loss of E-cadherin- relatively little is known about the role of HO-1 in dental mediated cell adhesion. Indeed, most of the E-cadherin field. repressors, including Snail, Twist, and SIP-1/ZEB-2, seem We previously reported that HO-1 in human pulp cells to fulfill this requirement, and they induce complete EMT was induced by inflammatory cytokines and may play a role when overexpressed in epithelial cell systems. in cytoprotection (J Endod 2006:32:39-43). Nitric oxide The Akt/PKB family of kinases is a downstream effector (NO) also induces the defense protein HO-1 in human pulp of phosphatidylinositol 3-kinase (PI3K) and is frequently cells. Furthermore, this cytoprotective function of HO-1 is activated in human cancers including OSCC. Recently, related to the activation of guanylate cyclase and the cGMP activation of the PI3K/Akt axis is emerging as a central pathway, suggesting a protective role for HO-1 in pulpal feature of EMT. Akt-induced EMT involves downregulation inflammation (Oral Surg Oral Med Oral Pathol Oral Radiol of E-cadherin, which appears to result from upregulation of Endod 2006;102:803-8). the transcription repressor Snail. However, the specific H2O2 has direct cytotoxic effect on human PDL cells, interactions of the Akt signaling with other E-cadherin which sequentially induces the defense protein HO-1 and repressors like Twist and SIP-1/ZEB-2 during EMT in up-regulates RANKL mRNA (J Perio Res 2007; IN Press). OSCC cells have not been clearly elucidated. Moreover, HO-1 not only plays a protective role against Akt activity is induced by ligand stimulation of growth H2O2-induced cytotoxicity in human pulp cells but also factor receptors, such as the insulin-like growth factor-I enhances odontoblast differentiation via the induction of receptor (IGF-IR) and the EGF family of receptors. Ligand DSPP (2007, Free Radical Biol Med, Submitted). Recently, stimulation activates PI3K, the upstream activator of Akt, by we reported that SP along with other modulators of neu­ direct binding to either the activated, phosphorylated roimmunoregulation to the induction of HO-1 and the receptor or to adaptor proteins phosphorylated by receptor inflammatory mediator MIP-3α/CCL20 in PDL cells, which kinase activity. Phosphoinositides generated by PI3K activity play an important role in the development of periodontitis or trigger activation of Akt kinases through direct binding to inflammation during orthodontic tooth movement. (Clin Exp the pleckstrin homology (PH) domain and the subsequent Immunology, 2007, in press). phosphorylation of Akt at two conserved residues. Therefore, HO-1 immunoreactivities were higher in oral lichen we used Akt inhibitor, structurally modified phosphati­ planus (OLP) mucosa than in normal mucosa; these results dylinositol ether lipid analogues (PIA), which specifically suggest that HO-1 plays a protective role for the damaged binds to the PH domain of Akt. epithelium of OLP lesions. Up-regulation of HO-1 in oral We have investigated whether treatment of Akt inhibitor epithelial dysplasias is also part of an early cytoprotection PIA could downregulate the Snail1, SIP-1/ZEB-2 and/or mechanism against carcinogenesis in the oral mucosa. In Oral Med Pathol 12 (2008) 61 oral cancer cells, NO-induced cytotoxicity was inhibited by disturbances; as the therapeutic management of them varies hemin and was enhanced in the presence of ZnPP IX, which depending on the actual disease progression, it is important was correlated with IRP1 and IRP2. HO-1 may play a major to differentiate them one another. However, both lesions role in mediating cytoprotection and iron homeostasis often show similar histological and radiological features, against NO toxicity in immortalized and malignant oral making it a diagnostic dilemma. In order to identify keratinocytes (2007, Cancer Letters, in press). important markers that permit differentiated diagnosis Collectively, HO-1 represents a cytoprotective enzyme among these disease entities, we performed immunological and, when expressed, produces therapeutic benefits in a and molecular analyses of OF, FD and OD with typical number of different conditions and diseases, such as pulpitis, histological and radiographic features. periodontitis, oral mucosal disease, and oral cancer. Stromal fibroblasts of FD produce bony matrix without morphologic evidence of osteoblastic cells while the bony 7. Identification ofmiR-31 and miR-211 as important matrix of OF and OD is surrounded by osteoblastic cells. miRNAs in oral carcinogenesis First, we therefore examined the difference among them in Chung-Ji Liu1-3, Shu-Chun Lin3*, Kuo-Wei Chang3* Runx2 expression (which determined osteogenic differen­ Department of Oral and Maxillofacial Surgery1, Taipei MacKay tiation from mesenchymal stem cells) because the difference Memorial Hospital, Taipei, Taiwan; MacKay Medicine2, of the Runx2 expression might permit the distinction among Nursing and Management College, Taipei, Taiwan; and Institute the three entities. As a result, fibroblastic cells of all three of Oral Biology3, School of Dentistry, National Yang-Ming diseases (FD, OF and OD) showed strong Runx2 expression University, Taipei, Taiwan in their nuclei. Second, we performed PCR analysis with MicroRNAs (miRNAs) are endogenous, short, non- peptide nucleic acid (PNA) for mutations at Arg201 codon coding RNA molecules involved in gene silencing and that of alpha subunit of the stimulatory G protein gene (GNAS), may be involved in tumorigenesis. This study aimed to which has reported to be a marker for extragnathic FD. All identify the miRNA expression profile and the functional cases with gnathic FD were positive for this mutation while roles of miRNA in oral squamous cell carcinoma (OSCC). none of OF and OD cases showed the mutation. These Tissue from 10 tumors and paired normal mucosa were used findings indicate that although these three are similar disease to quantitate the expression of a panel of 154 miRNAs using entities, especially in the demonstration of osteogenic stem-loop quantitative RT-PCR analysis, and 31 were found lineage in stromal fibroblastic cells. Furthermore, PCR to be either highly over- or under-expressed in the tumors, analysis with PNA for the GNAS mutations at Arg201 codon including 13 not previously reported to be dysregulated in is a useful method to differentiate FD from OF and OD. Our cancer. miR-31 and miR-211 were further analyzed in 40 data provide helpful information for the proper understanding, microdissected head and neck SCC specimens. miR-31 was classification and perhaps treatment of the fibro-osseous the most markedly up-regulated miRNA in primary OSCC, lesions. Furthermore, a number of other lesions demonstrate while that of miR-211 was down-regulated in OSCC tissues. clinical, radiographic and microscopic features similar to However, higher miR-211 expression was associated with those encountered in recognized fibro-osseous conditions. In vascular invasion, the most advanced nodal metastasis, and this lecture, other entities that may be included in the poor prognosis of OSCC. The expression and function of differential diagnosis of the fibro-osseous lesions also will lentivirus-transduced miR-31 and miR-211 was confirmed be discussed. by the stem-loop quantitative RT-PCR analysis and reporter assays showing repression of β-galactosidase through 9. Development of new strategies for the treatment binding of miRNAs to antisense sequences localized at the of oral cancer 3’-end of the gene. Expression of miR-31 and miR-211 Mark Yen-Ping Kuo significantly increased the growth, migration and anchorage- School of Dentistry, College of Medicine, National Taiwan independent growth of SAS OSCC cells and growth of University, Taipei, Taiwan tumor xenografts. These findings suggest that targeting of The chewing of areca nut (Areca catechu) preparations miR-31 may eventually be useful in OSCC therapy. The has long been associated with the high incidence of oral miRNA expression profile newly identified in the present submucous fibrosis and oral cancer in India and many study may contribute to future diagnostic and prognostic Southeast Asian countries. In Taiwan, there are two million studies of OSCC. people who have areca quid chewing habit. Oral cancer is the fourth leading cause of cancer death in the male population. 8. Molecular pathology of maxillofacial fibro-osseous Despite recent advances in surgery, radiotherapy and lesions chemotherapy, the survival of patients with oral cancer has Satoru Toyosawa not improved satisfactorily in past 30 years. Therefore, many Graduate School of Dentistry Osaka University, Osaka, Japan innovative approaches are being looked at for a cure. First, Maxillofacial fibro-osseous lesions, which characterized the local-regional biology of oral squamous cell carcinoma by replacement of by a fibrous connective tissue and (OSCC) makes this disease an ideal target for tumor mineralized-bony matrix, include ossifying fibroma (OF), suppressor gene delivery. Adenovirus-mediated p53 gene fibrous dysplasia (FD) and osseous dysplasia (OD), etc. therapy for cancer is currently undergoing phase I/II clinical They may be associated with cosmetic and functional trials in several countries. However, adenovirus-mediated 62 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology gene therapy has been reported to have acute toxicity and Nicotine enhances abnormal alterations in oral epithelium, immunological hazards. Recent studies have shown that which may induce periodontal diseases and oral cancer. The polyethylenimine (PEI) nonviral vector is significantly purpose of the present study was to investigate the effect of more potent in cells bearing mitotic activity. This higher nicotine on the expression pattern of hBD-1, -2 and -3 and efficiency of gene transfer using PEI in proliferative cells, the intracellular signaling pathway. The keratinocyte cell could reveal particularly interesting in heterogeneous tumor lines, HaCaT, were grown in DMEM supplemented with tissues, such as OSCC tissue, containing both proliferating 10% FBS. In order to examine the effect of nicotine on the tumor cells and differentiated non proliferating stromal expression of hBDs, the cells were incubated with 2.5, 5, 10 cells. Furthermore, PUMA (p53 upregulated modulator or 25 g/ml of nicotine (Sigma) for 4, 8, 12, 24 or 48 hrs. of apoptosis) was recently been found as an essential Expression of the hBD mRNAs was observed by RT-PCR mediator of p53-dependent and -independent apoptosis in and quantitative RT-PCR using TagMan probes (Applied vivo. Therefore, we used PEI nonviral vector to transfer Biosystem). Western blotting and ELISA assay (Peprotec) PUMA gene DNA for gene therapy on a murine oral cancer were performed to evaluate the level of hBD2 peptide. To xenograft model. We found animals treated with PEI/wild- elucidate the intracellular pathway in the expression, the type PUMA DNA have a significant reduction in tumor size cells were treated with inhibitors [SB203580(p38), when compared with those treated with PEI/mutant DNA MG132(NF-κB) or JNK2(JNK)] in a series of experiments. controls (P < 0.05). Our results suggest that PEI-mediated Luciferase reporter assay was performed for analysis of PUMA gene transfer is also a good candidate for adjuvant promoter activity. The statistical significance was analyzed therapy with radiation or chemotherapy. by One-way ANOVA. Expression of hBD-1 and -2 mRNA Secondly, phage display has recently been approved by was upregulated by nicotine in a dose- and time-dependent the FDA for use as therapeutics in a variety of diseases, manner. Incubation with nicotine at any dose did not change suggesting that the future looks bright for phage display in the expression level of hBD-3. The increased production of anti-cancer drug development. We identify several 12-mer hBD-2 peptide was confirmed by Western blotting and peptides specifically binding to a Taiwanese OSCC TW2.6 ELISA assay. Treatment with SB203580 or MG132 signifi­ cell line with phage display technique. The selected phages cantly inhibited the upregulated expression of hBD-2. When and corresponding peptides specifically bound to the cell the luciferase vectors with NF-κB mutant were transfected, surfaces of TW2.6 cells and a Taiwanese oral dysplasia cell the luciferase activity dramatically decreased. These results line TW1.5, but not normal oral keratinocyes. In SCID mice indicate that nicotine upregulates hBD-2 expression in bearing TW2.6 OSCC xenografts, the selected phages keratinocytes, possibly via p38MAP kinase and NF-κB specifically bound to the tumor and the effect was inhibited pathway. Increased production of hBD-2 may protect against by competition with corresponding peptides. Among all the infection when oral mucosa is exposed to nicotine. selected phage clones, C39-phage showed the highest targeting ability in tumor with 27.78-fold increases in 2. Uvula alterations associated with oral submucous concentration compared to control phage. Furthermore, fibrosis: a clinical study C39-peptide-linked liposomes that carried doxorubicin Der-Rong Lai1, Hui-Jen Chang1, Ying-Tai Jin2, Yeong- (C39-Lipo-Dox) not only suppressed tumor growth better Lei Huang3 but also showed no significant body weight change as Foo-Yin University Hospital1, Ping-Tung, Taiwan; National compared to Lipo-Dox treated groups. Kaplan-Meier Cheng Kung University2, Tainan, Taiwan; and Tai-Shin Dental analysis showed mice bearing TW2.6 OSCC xenografts Clinic3, Taipei, Taiwan with C39-Lipo-Dox treatment had significantly better overall (OSF) is characterized by a survival than those treated with Lipo-Dox and PBS. To our burning sensation in the oral cavity, blanching and stiffening knowledge, this is the first identification of a cancer-specific of the oral mucosa and oropharynx, and . Only peptide using the phage display approach in oral cancer and limited information can be obtained concerning uvula the novel peptide is a good candidate for targeted drug alterations associated with lesions of OSF. The purpose of delivery to OSCC. this study was to examine and classify the clinical pattern of destroyed uvula in patients with OSF. Eighty patients (68 men and 12 women; age range, 18-63 years; median 35.9 Oral presentation years) suffering from OSF diagnosed clinically and pathologically at the Foo-Yin University Hospital, Ping- 1. Nicotine upregulates β-defensin-2 expression Tung, Taiwan, were studied. A personal history of betel quid via p38MAP kinase and NF-κB pathway in kera­ chewing habit was obtained from each patient. A visual tinocyte examination of the destroyed uvula of each patient was Yoshihiro Abiko, Sumiko Nakamura, Mami Yamazaki, undertaken by a single clinician together with the same Michiko Nishimura, Masato Saitoh, Tohru Kaku photographic condition. Results indicated a correlation and Pathology, Health Sciences University of between the destroyed uvula and OSF. Eighty patients Hokkaido, Sapporo, Japan exhibited five clinical discernible changes of the destroyed Human β-defensins (hBDs) belongs to a group of uvula: pattern I (normal, 12%), pattern II (variable in size antimicrobial peptides expressed mainly in epithelial cells. and shape, 48%), pattern III (bud appearance, 26%), pattern Oral Med Pathol 12 (2008) 63

IV (bulla, 8%), and pattern V (spontaneous disappearance, 4. matrix protein 1 (DMP1) expression 6%). Data from this study indicate that various-degree in osteomalacia-associated tumor with different changes of the destroyed uvula are not in proportion to the histopathologic entities severity of the OSF. The high frequency of friction and Sunao Sato, Michiko Yuki, Mitsunobu Kishino, Yuzo longer duration of quid chewing are the two important Ogawa, Satoru Toyosawa factors that predispose the discernible changes of the Osaka University, Suita, Japan destroyed uvula. Oncogenic osteomalacia (OO) is a rare disease characterized by hypophosphatemic osteomalacia associated 3. Prevalence of tobacco related oral lesions in with tumors that are termed phosphaturic mesenchymal truck drivers and helpers in north-western Indian tumors (PMTs). It has been shown that PMTs overproduce region fibroblast growth factor-23 (FGF-23) that depletes total- Simarpreet Sandhu1, Jagpreet Sandhu2, Vattan Sharma3 body phosphate by reducing tubular reabsorption of G.I.D.S.R.1, Ferozepur, India; D.I.R.D.S.2, Faridkot, India; and phosphate. When PMT is discovered and removed, the renal Amritsar3, India phosphate wasting stops and the osteomalacia resolves. Prevalence studies are important to determine the oral Immunohistochemistry of dentin matrix protein 1 (DMP1) health status and treatment needs of a population. The and FGF-23 was a useful diagnostic tool for identifying objectives of the present study were (1) to determine the PMTs with typical histology (mixed connective tissue frequency of oral mucosa lesions and to analyze their variant). However, PMTs are usually lack specific histological relationship with toxic habits (tobacco and alcohol) in truck features and they show diverse histopathology. In fact, drivers; and (2) to establish necessities for treatment and despite their association with OO, there are many PMTs that opportune preventive measures. The study sample consisted are diagnosed as other mesenchymal tumors with different of 405 truck drivers and helpers in an exclusive health camp histopathologic entities. In this study, to verify whether co-organized by a leading oil company and a retail outlet DMP1 or FGF-23 is a useful marker for identifying PMTs “Hardas Fuel Point” where the participants frequented for with different histopathologic entities (giant cell tumor, fueling options. The clinical examinations were performed glomangiopericytoma, fibrosarcoma, and ossifying fasciitis), by both the authors. All participants were questioned about we performed immunohistochemistry of DMP1 and FGF-23 their general health status, medication use, oral hygiene in these PMTs, in addition to PMTs with typical histology. habits, tobacco and alcohol use, and education status. The All cases of PMTs clearly showed positive immunoreaction participants’ ages ranged from 18 to 62 years. The majority for DMP1. All of them also showed positive immunoreaction were North Indians (n = 378, 93.3%); of these 49% were for FGF-23 although their immunoreaction was not clearly from Punjab, 26% from Uttar Pradesh, and 25% from other visible. These findings indicate that DMP1 immunostaining neighboring states. The prevalence of smokers’ tobacco use is a more useful tool than FGF-23 immunostaining for was 49% (smoking 31%). Mean age of initiation was 21 identifying PMTs irrespective of their histopathology. years. Median deviation was 10 years. Many subjects reported using more than one type of tobacco and abuse of 5. Tenascin in different histological grades of oral sub­ alcohol. Oral leukoplakia was present in 49% of smokeless mucous fibrosis - an immunohistochemical appraisal tobacco users and 1.5% of non-users. Prevalence of oral Nirmala N. Rao1, Jalaj Tak1, M.S. Kotian2 leukoplakia increased with duration of chewing, frequency Department of Oral Pathology1, Manipal College of Dental of chewing per day, period of time chewed, and retention of Sciences, Manipal, India and Department of Community chewing quid overnight. Among subjects with oral leuko­ Medicine2, Kasturba Medical College, Mangalore plakia who underwent biopsy, 61 had benign hyperkeratosis, The distribution of extracellular matrix protein tenascin 4 mild dysplasia, 1 moderate dysplasia, and 2 severe was studied in 30 cases of oral submucous fibrosis (OSF) by dysplasia. Among tobacco abusers, the other lesions encoun­ employing the monoclonal antibody to tenascin. In normal tered were smokers’ melanosis (n = 50, 12.3%), nicotinic mucosa, the immunoreaction of tenascin was observed as a (n = 22, 5.43%), tobacco pouch keratosis (n = 38, continuous thin delicate line, restricted to basement 9.38%), and submucous fibrosis (n = 9, 2.23%). Overall membrane and around the blood vessels, with an enhanced prevalences of dental caries, , and plaque did not expression of tenascin in the basal cells of the oral differ between users and non-users. In conclusion, long epithelium. A similar mode of localization of tenascin was strenuous working hours and difficult terrains lead to a very observed in early stages of OSF with minimal extension into high psychological dependence. Special efforts are needed the underlying connective tissue. Areas of diffuse staining in to educate the truck drivers and helpers about the ill effects the deeper connective tissue were observed in early and of tobacco and alcohol consumption using available moderately advanced stages of OSF indicating an accentu­ modalities such as oral health exhibitions and camps. ation of its production by the mesenchymal cells especially in zones of fibrosis. The enhanced expression of tenascin suggests that tenascin plays a role in organizing and remodeling the stroma. Studies have shown that tenascin is a variable marker for identification of the early stage of OSF. However, the present study showed a distinct correlation 64 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology between tenascin expression and density of inflammatory such as osteonectin (ON), bone sialoprotein (BSP), osteo­ reaction particularly in early and moderately advanced pontin (OPN), osteocalcin (OC), and alkaline phosphatase stages of OSF. (ALP), in human pulp cells. H2O2 decreased cell viability, but increased HO-1 and DSPP expression in a concentration- and time-dependent manner, whereas the mRNA expression Poster presentation levels of OPN, OC, and ALP were decreased. H2O2 did not affect the mRNA expression levels of ON and BSP. We also 1. Defense mechanism of heme oxygenase-1 against found that H2O2-induced expression of mRNAs for HO-1 cytotoxic and receptor activator of NF-kB ligand and DSPP was enhanced by hemin and 8-bromo-cGMP, inducing effects of hydrogen peroxide in human while it was reduced by a selective inhibitor of guanylate periodontal ligament cells cyclase, 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ). Sung-Hee Pi, Sang-Cheol Kim, Hyun-Tae Kim, Hwa- Furthermore, inhibitors of phosphatidylinositol 3-kinase Jeong Lee, Suk-Keun Lee, Eun-Cheol Kim (PI3K), ERK, and p38 MAP kinase blocked H2O2-induced Department of Oral and Maxillofacial Pathology, Wonkwang cytotoxicity and the expression of HO-1 and DSPP mRNAs University, Iksan, South Korea in pulp cells. Treatment with N-acetyl-cysteine (NAC) and Although induction of heme oxygenase-1 (HO-1) by bilirubin attenuated H2O2-induced cytotoxicity and the H2O2 has been reported, the protective role of HO-1 expression of HO-1 and DSPP in pulp cells, while vitamin E against the cytotoxic and osteoclastogenic effects of H2O2 did not have these effects. These data suggest that the has not been elucidated in human periodontal ligament induction of HO-1 by H2O2 in pulp cells plays a protective (PDL) cells. The aim of this work was to investigate the role against the cytotoxic effects of H2O2 and stimulates defense mechanism of HO-1, on H2O2 induced cytotoxicity DSPP expression, resulting in premature odontoblast different­ and expression of receptor activator of NF-kB ligand iation through pathways that involve cGMP, PI3K, p38, and (RANKL) and osteoprotegerin (OPG) as markers for ERK. This study was supported by a grant of the Korea osteoclast differentiation in PDL cells. Using human PDL Health 21 R&D Project, Ministry of Health & Welfare, cells, the cytotoxicity was measured by MTT assay, Republic of Korea (A06-0005-AL1018-06N1-00010A). and expression of HO-1, RANKL and OPG mRNA was determined by reverse transcription-polymerase chain 3. Functional interaction between nitric oxide- reaction. H2O2 produced cytotoxic effect by reducing the cell induced iron homeostasis and heme oxygenase-1 in viability and enhancing the expression of HO-1 and RANKL immortalized and malignant oral keratinocytes mRNAs in a concentration- and time-dependent manner. Sun-Kyung Lee, Dae-Yeol Park, Hwa-Jeong Lee, Jun Additional experiments revealed that HO-1 inducer (hemin), Lee, Moon-Ki Choi, Byung-Hun Jeon, Chang-Duk membrane-permeable cGMP analog (8-bromo-cGMP), Jun, Suk-Keun Lee, Eun-Cheol Kim carbon monoxide,­ the ERK and p38 MAP kinases inhibitor, Department of Oral and Maxillofacial Pathology, Wonkwang protein kinase inhibitor (KT5823) and NF-kB inhibitor University, Iksan, South Korea (pyrrolidine dithiocarbamate), also blocked the effects of Heme oxygenase-1 (HO-1) is involved in a variety of H2O2 on cell viability and RANKL mRNA expression in regulatory and protective cellular mechanisms as a stress- PDL cells. These data suggest that HO-1 induction plays responsive protein. Whether HO-1 plays a protective role a protective role in PDL cells against the cytotoxic and against NO-induced cytotoxicity in oral cancer cells has not RANKL inducing effects of H2O2 through multiple signaling yet been established. We used sodium nitroprusside (SNP) pathways. This study was supported by a grant of the Korea as a source of exogenous NO in studies of NO-induced Health 21 R&D Project, Ministry of Health & Welfare, cytotoxicity in immortalized (IHOK) and malignant oral Republic of Korea (A06-0005-AL1018-06N1-00010A). keratinocytes (HN12). The roles of the caspase pathway, of regulatory proteins of iron metabolism (iron regulatory 2. The role of heme oxygenase-1 in the cytotoxic protein (IRP)1, IRP2, transferrin receptor (TfR), and and dentin sialophosphoprotein mRNA-inducing ferritin), and of HO-1 in protection against NO-induced effects of hydrogen peroxide in human pulp cells cytotoxicity were assessed. The SNP-induced growth Kyung-San Min, Hwa-Jeong Lee, Sung-Hee Pi, Hoon- inhibition and apoptosis of IHOK and HN12 cells was Sang Chang, Kyung-Hwa Kang, Hyung-Ryong Kim, reduced by addition of ferric citrate (FC). At low Hong-In Shin, Suk-Keun Lee, Eun-Cheol Kim concentrations (< 1 mM), SNP up-regulated cellular iron Department of Oral and Maxillofacial Pathology, Wonkwang metabolism by increasing expression of IRP1, IRP2, and University, Iksan, South Korea TfR, whereas at high concentrations (> 2 mM), SNP down- Although the induction of heme oxygenase-1 (HO-1) by regulated expression of these proteins. A consistent corre­ hydrogen peroxide (H2O2) has been reported, the protective lation between decreased levels of IRP1, IRP2, and TfR and role of HO-1 against the cytotoxic effects of H2O2 has not increased NO-induced cytotoxicity and apoptosis was been clarified in human pulp cells. In this study, we observed. Addition of FC inhibited the NO-induced decrease investigated whether HO-1 is involved in H2O2-induced in IRP1, IRP2, and TfR levels. Moreover, SNP increased the cytotoxicity and examined the production of dentin sialo­ expression of HO-1 and ferritin in IHOK and HN12 cells in phosphoprotein (DSPP) and other mineralization markers, a concentration-dependent manner. NO-induced cytotoxicity Oral Med Pathol 12 (2008) 65 was also inhibited by hemin (an HO-1 agonist) and was efficacy of DNA vaccines. In this study, we used this HOPV enhanced by zinc protoporphyrin IX (an HO-1 inhibitor). hamster model to compare the anti-tumor effectiveness of Based on these results, we conclude that HO-1 plays a major different procedures of DNA vaccine delivery, intramuscular role in mediating cytoprotection and iron homeostasis injection alone or with electroporation (EP). Forty hamsters against NO toxicity in immortalized and malignant oral were divided equally into four groups. These groups were keratinocytes. designated as group-N (no treatment), group-V (Vaccination without EP), group-E (EP without vaccination), and 4. Expression of heat shock proteins in rat oral group-VE (vaccination followed by EP). The animals in mucosa after CO2 laser irradiation group-N were injected intramuscularly with vector only, Akira Yamasaki, Hiroshi Ito, Yuuko Sakurai, Noriko while those in group-V were injected with DNA plasmids Okuyama encoding the E6 gene. The animals of group-VE were Department of Oral Medical Sciences, Oral Pathology, Ohu injected DNA plasmids followed by EP, while those in University School of Dentistry, Koriyama, Japan group-E were only injected vector followed by EP. Three The CO2 laser is now widely used in a variety of surgical weeks after the initial DNA injection, all animals were procedures and has been proven to be a useful device, but the boosted with the same plasmids that they had received tissue responses to the irradiation is not fully understood. In initially. The lingual tips of hamsters were painted three this study we examined the expression of heat shock times a week with DMBA for 8 weeks. The middle portion proteins, Hsp27, Hsp47, and Hsp70, to elucidate the tissue of the lingual tip was then excised under anesthesia. responses of rat oral mucosa to CO2 laser irradiation. Pulsed Thereafter, the tips were painted daily with DMBA until the CO2 irradiation was performed on the palatal mucosa of animals were sacrificed. All hamsters of N and E groups male Wistar rats, 7-9 weeks of age, with the parameters of showed lingual carcinoma. Some delays in cancer 600 μs pulse duration, 5 W peak power, 151.52 Hz repetition development in the hamsters of V and VE group were rate, and 4.72 J/mm2 energy density. At 30 min, 6, 12, 24, 72 observed. In particular, in VE group, 6 hamsters showed no hrs after irradiation, the palatal mucosa was dissected and lesions. These results suggest that immunization with E6 examined by immunohistochemistry. The CO2 laser DNA vaccines followed by EP in vivo delays development irradiation on the palatal mucosa created a bowl-shaped of papillomavirus-associated oral carcinoma. In vivo EP is a necrotic area including lamina propria 6 hrs after irradiation, potent method for DNA vaccine delivery. a strong expression of Hsp70 was observed in both epithelial cells and connective tissue cells directly facing the necrotic 6. A histopathological and immunohistochemical tissue, the expression being reduced after 72 hrs. study of cell adherence in pleomorphic adenoma Constitutively found in the spindle cells adjacent to the Hisae Anzai, Tadahiko Utsunomiya, Hirotsugu Yamamoto epithelium, the expression of Hsp47 was enhanced 24 hrs Department of Oral Pathology, Nihon University School of after irradiation and thereafter increased coincident with the Dentistry at Matsudo, Chiba, Japan progress of repair process. After laser irradiation, basal and Pleomorphic adenoma is the major histopathological suprabasal layers of palatal epithelium exhibited the type of salivary gland tumors. Histologically, the tumor expression of Hsp47, as well. Although a strong expression consists of epithelial and mesenchyme-like elements of Hsp27 was constitutively detected in the whole layers of indicating “mixed appearance”. Because the periphery of the palatal epithelium, vascular endothelial cells, and smooth epithelial elements shows loss or weakness of cell adherence muscle cell, no significant changes were discernible in the and continues to myxomatous tissues, it is thought that cell laser wound throughout the experiment. From the present adherence play an important role to form the “mixed study, it is suggested that strongly expressed in response to appearance”. This study evaluated the expression of E- and CO2 laser irradiation is Hsp70, whereas Hsp47 and Hsp27 N-cadherin in order to clarify the histopathological nature of involve mainly in repair process through their function as cell adherence in pleomorphic adenoma. We found that the molecular chaperons. center of the epithelial elements with ductal, cord-like or sheet-like proliferation showed positive immunoreactivity 5. Electroporation improves the efficacy of DNA mainly for E-cadherin and partly for N-cadherin. Periphery vaccines against hamster oral papillomavirus- of epithelial elements and mesenchymal-like tissue indicated associated oral cancer positive immunoreactivity for N-cadherin. Our results Hatsuhiko Maeda, Shinichi Komatsu, Takashi Umebayashi, suggest that the changes of expression of E- and N-cadherin Katsutoshi Kubo, Yoshihiko Sugita, Emiko Sato are associated with epithelial-to-mesenchymal differentiation Aichi-Gakuin University, Nagoya, Japan in pleomorphic adenoma. In a previous investigation, we developed a highly reproducible carcinogenesis model by combining DMBA application with physical wounding of the hamster lingual mucosa. Using this animal model, we demonstrated the presence of a novel hamster oral papillomavirus (HOPV). Recently it has become clear that more potent methods for DNA vaccine delivery need to be developed to enhance the 66 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology

7. Elevated expression of ERK 2 in adenoid cystic exhibited cellular and nuclear atypia with occasional carcinoma treated with PD98059 mitoses. The more significant proliferative activity than in Ji-an Hu, Yining Li, Songying Li PA was also confirmed by the occasional presence of Ki-67 Department of Oral Pathology, School of Stomatology, Zhejiang positive cells. Two cases showed overexpression of p53 and University, Hangzhou, China one case reduced expression of p27. There were no obvious Adenoid cystic carcinoma (ACC) is a familiar malignant pre-existing PA and cancerous and/or sarcomatous com­ tumor of the salivary gland. We examined the effect of ponents. It may be appropriate at present to separate these chronic exposure of tumor cells to a mitogen-activated tumors from ordinary PA, carcinoma ex PA and carcino­ protein kinase/extracellular signal-regulated kinases (ERK) sarcoma by naming them “low-grade malignant PA”. kinase inhibitor, PD98059, on cell proliferation. ACC cells Although the histopathological findings indicate low-grade were cultured in the presence of PD98059 for more than 4 malignancy, it is necessary to analyze a large number of weeks and proliferation and signal transduction pathways cases with long-term follow-up in order to evaluate the were examined. PD98059 significantly inhibited the precise prognosis. Recognition of the tumors with above- proliferation of parental cells. PD98059 failed to inhibit mentioned characteristics may also be necessary to avoid the proliferation of ACC-M cells significantly. Expression of misdiagnosis, because of their resemblance to other ERK 1 and 2 was elevated in these cells. These phenotypes epithelial-myoepithelial tumors such as epithelial-myo­ were reversible. Downregulation of ERK 2, but not ERK 1, epithelial carcinoma and basal cell adenocarcinoma. by small interfering RNA significantly inhibited the proliferation of ACC-M cells. Taken together, chronic 9. COX2 expression in oral squamous epithelium exposure of tumor cells to PD98059 induced elevated may relate to prickle cell differentiation expression of ERK 2, which was associated with decreased Kazuo Komiyama1,3, Yukako Miki2, Koichi Nagai1, sensitivity of cellular proliferation to PD98059. Treatment Hidero Oki2 of patients with a MEK inhibitor in combination with Department of Pathology1, Department of Oral and Maxillofacial inhibitors of protein synthesis or translation could be an Surgery2, and Dental Research Center3, Nihon University important option for future clinical studies. School of Dentistry, Tokyo, Japan COX2 is a key enzyme to produce PGE2 from arachidonic 8. Pleomorphic adenoma with atypical histopatho­ acid. Overexpression of COX2 was identified in many logical features suggestive of low-grade malignancy malignancies including oral cancer. Previous papers reported - Low-grade malignant pleomorphic adenoma that high COX2 expression ratio of cancer cells candidates Ikuko Ogawa, Mohamed Mahmoud Abdelrazk, Yasusei as a prognostic factor of the oral cancer. In the present study, Kudo, Mutsumi Miyauchi, Yoshiya Shimao, Takashi we examined coordinate expression of sPLA2-X and COX2 Takata in oral mucosal epithelium with precancerous condition (30 Hiroshima University and Miyazaki Prefectural Hospital, cases) and cancer (30 cases) as well as normal epithelium Hiroshima, Japan (10 cases), focused on the structure of squamous epithelium Pleomorphic adenoma (PA) is the most common benign and prickle cells differentiation. We performed an in situ tumor of salivary glands. According to the present WHO hybridization and immunohistochemistry to detect these classification, carcinoma ex PA, carcinosarcoma and enzymes on the specimens, fixed in 10% neutral buffered metastasizing PA are classified as a malignant counterpart of formalin and paraffin embedded. In addition, the role of PA. In this study, we report four cases of which histological COX2 and sPLA2 was examined in cell proliferation using and immunohistochemical features are essentially identical human oral cancer cells. Inhibitors, NS398 for COX2 and to those of PA nevertheless accompanying more or less 4-BPB for sPLA2 were added to the oral cancer cell culture, malignant characters such as infiltrative growth and/or cell respectively. MMT assay was employed for counting cell atypia. They could not fit into the diagnostic criteria of the numbers. Normal epithelium showed minimum level or present WHO classification. Therefore, we propose a new no expression of both enzymes. Precancerous conditions entity of malignant counterpart of PA, “low-grade malignant including oral and dysplasia showed varied PA”. All four cases arose in palatal gland and macroscopically degrees of COX2 expression, which strongly correlated were well circumscribed, but microscopically somewhat with the presence of prickle cell layer. In oral cancer, high infiltrative in three of them. The tumors were composed of expression cells were also found in well-differentiated type of glandular epithelial and neoplastic myoepithelial cells squamous cell carcinoma but not in the undifferentiated type (NMCs) arranged in tubular, trabecular and solid patterns. and basal cell carcinoma. In vitro study clearly showed that Immunohistochemically, glandular cells were positive for the COX2 and sPLA2 maintain the cell number of cultured EMA and NMCs exhibited various degrees of positive oral cancer cells. These results showed that COX2 synthesis reaction for myoepithelial markers such as smooth muscle was related to the prickle cell differentiation in squamous actin, S-100 protein and GFAP, similar to the immunoreactivity cell epithelium or in oral cancer. These phenomena indicate of PA. Hyalinized stroma was prominent with some myxoid that elimination of cell apoptosis resulted in thickening of areas. Although chondroid areas were not observed, NMCs the epithelium and tumor cell growth. focally showed isolation into the hyaline or myxoid matrices. The remarkable feature was that the tumor cells diffusely Oral Med Pathol 12 (2008) 67

10. Low-grade papillary cystadenocarcinoma of high expression of HOX C13. The expression of HOX C13 the posterior floor of the mouth first manifested as mRNA in odontogenic lesions had heterogeneity. The a metastatic neck node with cytology mimicking a expression of HOX C13 could improve the epithelial thyroid papillary carcinoma proliferation, and loss of expression may lead to the Julia Yu Fong Chang1, Shirley Yan2, David Watkins2, cornification and degeneration of epithelial cells. Harvey P Kessler3 Department of Diagnostic Sciences1, Baylor College of Den­ 12. Metastatic thyroid cancer incidentally found tistry, Texas A&M Health Science Center; School of Dentistry in lymph nodes from neck dissection material of and Dental Department of National Taiwan University Hospital, buccal cancer College of Medicine, National Taiwan University, Taipei, Naoyuki Matsumoto, Syotaro Mukae, Yusuke Amano, Taiwan; Department of Pathology2, Baylor University Medical Kazuo Komiyama Center at Dallas; and Department of Diagnostic Sciences3, Department of Pathology, School of Dentistry, Nihon University, Baylor College of Dentistry, Texas A&M Health Science Center, Tokyo, Japan Dallas, USA Asymptomatic cancer is rarely found during the course Tumors with first manifestation as a metastatic neck of histopathological examination. We reported a case of node are commonly seen in nasopharyngeal carcinoma and small primary thyroid carcinoma, incidentally found in the thyroid papillary carcinoma. Here, we reported a case of lymph node metastases from total neck dissection for buccal low-grade papillary cystadenocarcinoma of the posterior squamous cell carcinoma (SCC). A 61-year-old male floor of the mouth first presented as a metastatic neck node. referred to our hospital due to discomfort and a nodule at the Because the lesion showed a papillary growth and a bland buccal mucosa. The patient recognized the nodule a month looking cytology with nuclear overlapping, nuclear grooving, before. The nodule was gradually increasing in size. Biopsy and ground-glass nuclei mimicking a thyroid papillary specimen showed a moderately-differentiated SCC (T3N2bM0). carcinoma, a total thyroidectomy was performed. However, The tumor was excised and radical neck dissection was only one occult thyroid papillary carcinoma focus was performed simultaneously. Seventeen lymph nodes were identified. Nine months later, a tumor located in the posterior obtained and metastasis of SCC was found in a submandibular floor of the mouth which was further proved pathologically lymph node (n = 1). Moreover, metastases of papillary as a low-grade papillary cystadenocarcinoma was found. carcinoma were found in deep lateral cervical lymph nodes The tumor was then excised. Both the metastatic tumor in (n = 6), suggestive of thyroid cancer. The radiological the neck node and the tumor at the posterior floor of the examination (CT, MRI, US) including the Tl-Tc scintigraphy mouth shared similar histologic features and were TTF-1 failed to detect the thyroid cancer. Only a vocal fold paralysis negative. Although a metastatic papillary carcinoma in the was found by the fiberscope examination, suggestive of the neck node most commonly comes from the primary lesion of invasion of the thyroid carcinoma to recurrent laryngeal a thyroid papillary carcinoma, it may occasionally arise nerve. The patient was undergone hemi-thyroidectomy and from other primary oral papillary carcinomas. A routine cervical dissection. There were small foci (< 4 mm) of immunohistochemical study of TTF-1 for a neck node well-differentiated papillary carcinoma with lymphatic before thyroidectomy is helpful to avoid misdiagnosis of a vessel invasions and marked fibrous stroma. Metastasis was metastatic papillary carcinoma from primary sites other than also found in a pre-tracheal lymph node. We have the thyroid. demonstrated an unusual case of double cancer, detected only by a histopathological examination. Follow-up CT 11. Expression of HOX C13 in odontogenic lesions examinations have been performed for 6 years and show no Ming Zhong1, Jie Wang1, Jie Liu1, Bo Zhang2 evidence of cancer recurrence. China Medical University1, Shenyang, Liaoning, P. R. China and Peking University2, Beijing, P. R. China 13. Expression of type IV collagen in different This study assessed the expression of HOX C13 mRNA histological grades of oral squamous cell carcinoma in odontogenic lesions. HOX C13 mRNA levels were - an immunohistochemical study examined in 47 cases of ameloblastoma (AB), 3 cases of Pankaj Agarwal, Nirmala N Rao calcifying cystic (CCOT), 3 cases of Manipal College of Dental Sciences, Manipal, Karnataka, ameloblastic fibroma (AF), and 2 cases of calcifying India epithelial odontogenic tumor (CEOT) by in situ hybridization. The purpose of this study was to evaluate the expression Ten cases of keratocystic odontogenic tumor (KCOT), and 7 of type IV collagen in normal oral mucosa (NOM) and cases of normal oral mucosa were selected as controls. HOX various histological grades of oral squamous cell carcinoma C13 mRNA was expressed positively in all odontogenic (OSCC), in order to determine whether this protein can be epithelium except AF. The positive rates were 97.9% (46/47) used as a marker in early detection of the biological behavior in AB, 100% (3/3) in CCOT, 100% (2/2) in CEOT, 70.0% of phenotypically altered cells. This study used an (7/10) in KCOT, and 42.9% (3/7) in the normal oral mucosa. immunohistochemical technique to examine the expression There was a significant difference among AB, KCOT and of type IV collagen in 10 specimens of histologically well- normal mucosa (x2 = 21.665, P < 0.001). The genesis and differentiated SCC (WDSCC), 10 specimens of moderately- development of the odontogenic tumors are related to the differentiated SCC (MDSCC), 10 specimens of poorly- 68 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology differentiated OSCC (PDSCC), and 5 specimens of NOM as cystic carcinoma cells. controls. Staining intensity, staining pattern, and distribution of type IV collagen were interpreted in different histological 15. Immunohisotchemical analysis of activated grades of OSCC and NOM, and were compared among den­dritic cells in oral lichen planus groups. Chi-square test showed highly significant differences Shotaro Mukae1, Yasuhiro Okaue2, Hidero Oki2, Kazuo in intensity of staining (P = 0.004), distribution (P = 0.003), Komiyama1 and pattern of staining (P = 0.001) of type IV collagen. Department of Pathology1 and Department of Oral and Fisher’s exact test also revealed statistically significant Maxillofacial Surgery2, Nihon University School of Dentistry, differences in inter-group relation for intensity of staining of Tokyo, Japan type IV collagen between WDSCC and PDSCC (P = 0.005), Oral lichen planus (OLP) is a chronic inflammatory and for distribution of type IV collagen between WDSCC lesion with unknown etiology. The lesion was characterized and MDSCC (P = 0.003). There was also a significant by an intraepithelial and subepithelial infiltrate of CD4 and difference in pattern of staining of type IV collagen between CD8 positive cells together with Langerhans/dendritic cells WDSCC and PDSCC (P = 0.001). We conclude that type IV (LC/DC). The purpose of this study was to determine the collagen decreases from WDSCC to PDSCC, showing a activation level of LC/DC in the lesion of OLP and normal direct relationship with the differentiation of the tumor cell. oral mucosa (NOM) by immunohistochemical methods. Also, the intensity of staining of type IV collagen decreases Biopsies from 30 patients diagnosed clinically and from WDSCC to PDSCC, indicating the decreased capacity histopathologically as suffering from reticular LP of the of undifferentiated epithelial cells to synthesize basement buccal and gingival mucosa were used. Serial sections were membrane components including type IV collagen. prepared and stained with Langerin, Fascin, s-100s, CD4, CD8, CD57, CCR7, D2-40, and COX-2 antibodies using 14. FGF7/KGF enhances cell proliferation but DAKO CSAII kit. LC/DCs found in the epithelium and inhibits cell invasion of adenoid cystic carcinoma submucosa of the OLP lesions and NOM were counted cells positive for the Langerin, Fascin, s-100s and CCR7. LC/ Satoshi Maruyama, Jun Cheng, Takashi Saku DCs were reliably identified using Langerin and s-100s Divisions of Oral Pathology, Niigata University Graduate antibodies in samples of NOM. The number of Langerin- School of Medical and Dental Sciences, Niigata, Japan positive cells was significantly increased in the lesional Our microarray analysis by using cancer-associated epithelium of OLP (P = 0.003) compared with normal oral DNA chips showed higher expression levels of fibroblast epithelium. Fascin and CCR7 positive cells were not found growth factor 7 (FGF7/KGF) in ACCM, a highly metastatic in normal oral epithelium, while in lesional epithelium of clone derived from ACC2, a human salivary gland adenoid OLP the positive cells clustered in capillary of submucosa. cystic carcinoma cell system. We wanted to determine how In addition, activated LC/DCs migrating into the lymphatic FGF7 functions in metastatic steps through the FGF capillary were detected using double staining methods of receptors-signaling pathway. Total cellular RNAs and Fascin and D2-40. Overexpression of COX-2 was found in proteins were extracted from ACC2 and ACCM cells. Their prickle cells and recruited CD4 and CD8 positive cells in the mRNAs were purified and subjected for RT-PCR analyses, OLP lesions. The results of this study showed an increase of and their cell lysates were used for WB analysis. Cultured Langerin and Fascin-positive cells in OLP and indicated that cells on chamber slides were fixed with 4% paraformaldehyde these cells are immunologically active. COX2 overexpression for immunofluorescence for FGF7 and FGF receptors indicates that local PGE2 synthesis is probably involved in (FGFRs). For cell growth analyses, cells were seeded at a final activation of intraepithelial LC/DCs. Further, activated cell concentration of 0.4 x 105 each onto 35 mm dishes and LC/DCs migrate to regional lymph nodes. Taken together, cell numbers were counted daily for 7 days. For cell the antigen in OLP lesion development remains uncertain; invasion, cells were seeded on cell culture insert chambers however, DCs play a role in presentation to autoreactive coated with matrigel in the presence and absence of FGF7. cytotoxic T cells in the OLP lesion. ACCM cells showed higher expression of FGF7 at both gene and protein levels than ACC2. ACC2 and ACCM 16. Measurement of the Rontgen photo density of expressed FGFR1/4 genes but no FGF2/IIIb, which had the recovery process of the periapical abscess after been considered to be specific to FGF7. ACCM showed treatment of the necrotic pulp two-fold cell numbers of ACC2 on days 4 to 6. When Suhardjo Sitam recombinant FGF7 was added to the medium, ACC2 started Department of Dentomaxillofacial Radiology, Faculty of to proliferate as much as ACCM did. On the other hand, Dentistry, University of Padjadjaran, Bandung, West Jawa, ACC2 showed more invasive activity than ACCM, but its Indonesia invasive activities were suppressed in the presence of The purpose of this study was to know the recovery recombinant FGF7. FGF7/KGF, expressed in highly process of the periapical abscess after treatment of the metastatic ACCM, was shown to enhance its proliferation necrotic pulp by measuring the Rontgen photo density. We but suppress its invasion through FGFRs1/4 by autocrine analyzed the three studies related to recovery process of manners. These seemingly ambivalent tendencies in the the periapical abscess after treatment of the necrotic pulp cellular activity may benefit metastatic events of adenoid using as many as 90 Rontgen photo archives, which were Oral Med Pathol 12 (2008) 69 then measured by densitometer. The three studies using while CK13 was not positive with exceptional cases of the Student statistic test showed that the F count was more verrucous type (13 of 64 foci, 19%). All of the SCC foci (26 than the F table, which had significant meaning that showed of 26 foci, 100%) were positive for CK17 but not for CK13. the increase of the density value of the Rontgen photo files It was thus contrastive between CK17 positive areas and after treatment of the necrotic pulp. We conclude that the CK13 not-positive areas. Simultaneous positivities for photo density of the recovery process of periapical abscess CK13 and CK17 were obtained only in SCC (1 focus, 4%) after treatment of the necrotic pulp can be measured by and CIS (10 foci, 15%), both of which had distinct densitometer. keratinization, but not in other categories. The results indicate that this combined immunohistochemistry for these 17. Histopathological changes of low-energy laser two keratin species is a valuable aid for objective diagnosis irradiation on wound healing of exposed dental of oral borderline malignancies. pulp tissues Tadahiko Utsunomiya, Hirotsugu Yamamoto 19. The relationship between the state of oral Nihon University School of Dentistry at Matsudo, Chiba, hygiene and human papillomavirus infection Japan Yoshie Nishimura, Hatsuhiko Maeda, Keiko Toyama, The purpose of this study was to investigate the effects Haruo Nakagaki, Masami Hattori, Shinichi Komatsu, of low-energy laser irradiation on wounded pulp tissues in Takashi Umebayashi, Katsutoshi Kubo, Yoshihiko dogs. Gallium-aluminum-arsenide diode laser (300 mW) Sugita, Emiko Sato irradiation was applied to the pin-pointed-exposed surface Aichi-Gakuin University, Nagoya, Japan of the pulp (irradiation group), and histopathological Recently, some investigators described that human changes were observed at 1, 3, and 7 weeks after operation papillomavirus (HPV) infection was related to some factors compared to the changes of non-laser irradiation (control) including sex, age, occupation, smoking habit, alcohol group. Although degeneration, necrosis, capillary proliferation/ consumption, denture wearing, and others. The purpose of dilatation, and inflammatory infiltration were observed in this study was to investigate the correlation between changes both irradiation and control groups, fibroblastic proliferation in the state of oral hygiene and HPV infection of high school and fibrous matrix formation were found in irradiation group students. In this study, 100 (56 males and 44 females) 1 week after operation. In addition, the capillaries and students were randomly selected from the students in odontoblastic cells with nuclear polarity were markedly Nisshin Aichi Prefectural High School. In a 3-year period migrated around and/or under the fibrous matrices. Calcified from 2004 to 2006, the state of oral hygiene was examined dentin bridge formation was already completed 3 weeks during routine teeth examination in two periodical heath after operation in the irradiation group, although the checks. According to change in the state of oral hygiene, the complete formation of calcified dentin bridge was also students were divided into three groups. These groups were observed 7 weeks after operation in the control group. These designated as group-W (worsened state in year 3 than year results suggest that low-energy laser irradiation accelerates 1), group-I (improved state in year 3 than year 1), and wound healing of the exposed pulp tissues. The present group-N (no change student group). Oral squamous cells study was supported by Nihon University Individual were collected from swabs of the buccal mucosa. For this Research for (05-115) (2005). procedure, informed consent was obtained. The rate of HPV infection in these 3 groups was examined by PCR. In the 18. Immunohistochemistry for CK17 in the differen­ group-W, the HPV positive rate was 38.5% and 100.0% in tial diagnosis of oral borderline malignancies year 1 and 3, respectively. The HPV positive rate of group-I Toshihiko Mikami1,2, Jun Cheng1, Akinori Funayama1,2, was 53.1% and 15.6% in year 1 and 3, respectively. In the Satoshi Maruyama1, Hiroko Ida-Yonemochi1, Susumu group-N, the HPV positive rate was 10.9% and 30.9% in Shingaki2, Chikara Saito2, Takashi Saku 1 year 1 and 3, respectively. The results of the present Divisions of Oral Pathology1 and Reconstructive Surgery for investigation indicate that an improvement in the state of Oral and Maxillofacial Region2, Niigata University Graduate oral hygiene decreases the rate of HPV infection and School of Medical and Dental Sciences, Niigata, Japan demonstrates that oral care may be an effective way to To evaluate the usefulness of immunohistochemistry for prevent the HPV infection-related oral lesions, including the cytokeratin (CK) species in the diagnosis of oral borderline oral squamous cell carcinoma. malignancies, we compared CK17/CK13 expression patterns in those lesion foci categorized by referring to the Ki-67 20. Immunohistochemical expression of thymidylate immunohistochemical mode. There was no positivity for synthase in oral squamous cell carcinoma CK17 (0 of 20 foci, 0%) but positive for CK13 (20 of 20 Yi-Ru Fang, Masahiro Wato, Kazuya Masuno, foci, 100%) in normal epithelia. Mild epithelial dysplasias Junichiro Kotani, Akio Tanaka showed same tendencies (CK17 & CK13, 0%). In moderate Osaka Dental University, Osaka, Japan dysplasias, only one focus was positive for CK17 (1 of 20 We evaluated the frequency of cells in the S phase of the foci, 5%), while CK13 was positive in most of them (18 of cell cycle in formalin-fixed, paraffin-embedded sections of 20 foci, 90%). Most of the CIS foci (64 of 68 foci, 94%) had 29 specimens of oral well-differentiated squamous cell positivities for CK17 in the prickle cell to keratinized layers, carcinoma (OWSCC) from 29 patients who had not received 70 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology medication or radiation therapy. Ki-67, a marker of the G1 to demonstrated a deposition of this antigen at the level M phases, was also examined for comparison with S-phase of basement membrane. In conclusion, the tumor cells cell frequency. The sections were mounted on silane-coated exhibited an altered synthesis and secretion of type VII slides, deparaffinized in HemoDe®, and retrieved in 10 mM collagen protein. Further as the degree of de-differentiation citrate buffer at pH 9.0 by using an autoclave at 121°C for 15 in tumor cells increased there was an observable lack of min. They were then immersed in 3% H2O2 to block secretion of the protein extracellularly. On the other hand, endogenous peroxidase activity and incubated with there is a failure to reassemble the component of basement thymidylate synthase (TS), a marker of S-phase cells, or membrane around these tumor cells. Ki-67 monoclonal antibodies for 60 min at room temperature. The sections were reacted with EnVision+ system for 30 min 22. Characterization of with immunohisto­ at room temperature and visualized with 3,3’ diamin­ chemical analysis obenzidine. Digital images of these stained sections were Michiko Yuki, Sunao Sato, Mitsunobu Kishino, Yuzo taken in mid-power fields (using a 20-power objective lens) Ogawa, Satoru Toyosawa using a Nikon CoolScope®, and the ratios of TS-positive and Department of Oral Pathology, Osaka University Graduate Ki-67-positive cells to all tumor cells examined were School of Dentistry, Osaka, Japan separately estimated by using Win ROOF image system®. The clinical term epulis has been used to describe TS positivity was observed in the nuclei and cytoplasm of localized and is thought to correspond tumor cells from the outermost areas to the inner areas of to the term “irritation fibroma” of the extra-gingival areas. tumor nests (mean percent TS-positive cells, 29.72% ± Epulis is thought to be non-neoplastic reactive lesions 29.57%). Ki-67-positive reaction was discernible in the derived from periodontal origin. On the basis of histologic nuclei of cells from the outermost areas to the inner areas of appearance, epulis can be classified into several types; tumor nests (mean percent Ki-67-positive cells, 49.06% ± fibrous, granulomatous, ossifying etc. Thus, the morpho­ 21.98%). OWSCC tumor cells (about 30%) are in S phase, logical characteristic of the epulis is well known, but there and remaining cells (about 20%) are in G1, G2, or M phase. were few reports of their pathogenesis. As mentioned above, In conclusion, it is suggested that tumor cells in S phase epulis is thought to be periodontal origin. Epulis often has outnumber those in G1, G2 or M phase in OWSCC. the formation of metaplastic bone while irritation fibroma doesn’t have. Recently, periodontal ligament has been 21. Expression of the anchoring fibril component reported to be composed of osteogenic cells, which express type VII collagen in different grades of oral squamous transcriptional factor Runx2. Therefore, we hypothesize that cell carcinoma - an immunohistochemical study epulis may also be composed of Runx2-expressing osteogenic Sudhir1, Nirmala N Rao1, M S Kotian2 cells. In this study, to elucidate their pathogenesis, we Manipal College of Dental Science1, Manipal, India and performed immunohistochemistry of Runx2 in both epulis Kasturba Medical College2, Mangalore (fibrous, granulomatous, ossifying types) and irritation Basement membranes are involved in a number fibroma. As a result of immunohistochemical analysis, many of biological processes and act as a three dimensional Runx2-positive osteogenic cells were detected in all ten supporting scaffold. They form an important barrier against cases of epulis, in addition to one case of ossifying epulis as tumor cell invasion. Recently, biochemical characterization positive control. However, Runx2-positive osteogenic cells and production of antibodies against different constituents were not detected in all ten cases of irritation fibroma of the of basement membrane have proved to be more reliable in extra-gingival areas. These results suggest that Runx2- tumor invasion and metastasis. The present study elucidates positive osteogenic cells exist in epulis derived from the the role of type VII collagen, a principle constituent of periodontal ligament and that epulis and irritation fibroma anchoring fibril links the basal lamina of oral mucosa to are different disease entities in the demonstration of the the underlying connective tissue, in oral squamous cell osteogenic lineage, although both lesions show similar carcinoma (OSCC) samples. We also observed the intensity histopathology. and location of type VII collagen immunohistochemically, and to consider this marker as a useful adjunct in grading 23. Relation of stathmin expression with tumor and to identify more potentially aggressive and invasive invasion in human tongue cancer margins in the lesion. Forty-two samples of OSCC and seven Hiroto Itoh1, Yoshinori Jinbu1, Yuri Obi1, Yukio controls from buccal mucosa were stained with mouse anti- Ooyatsu1, Yasuhiro Ueno1, Mikio Kusama1, Keiichi human collagen type VII, at a dilution of 1: 1000, visualized Tsukinoki2 by DAB substrate. The antigen-positive areas in relation to Jichi Medical University1, Tochigi, Japan and Kanagawa pattern and intensity of staining were then assessed in all the Dental College2, Kanagawa, Japan samples. The deposition of type VII collagen antigen was Stathmin is a cytosolic phosphoprotein expressed at high evident around and within the malignant cells. Statistical levels in most human cancer cells. Stathmin expression and evaluation of data by using Chi-square test and Fischer’s phosphorylation are linked to the control of cell differentiation exact test revealed very highly significant relationship in and proliferation. However, few studies of stathmin localization and intensity of staining of this antigen and expression in human oral squamous cell carcinoma have histopathological grades of OSCC. The control tissues been reported. We performed immunohistochemical staining Oral Med Pathol 12 (2008) 71 of stathmin and analyzed the relation with the mode of cytometric and DNA fragmentation assay in SNP-treated invasion in human tongue cancer. Oral biopsy specimens of group, but it was reduced in 590-nm irradiation group. NO, 42 cases of tongue cancer (from 30 men and 12 women) ROS, and ONOO− levels were increased after SNP treatment were used. Paraffin sections were deparaffinized and primary but ROS and ONOO− levels were decreased by 590-nm antibody (anti-stathmin monoclonal antibody) was added. irradiation, in spite of high NO concentration by SNP. The mode of invasion was divided into five types from Cytochrome C release was found in SNP-treated SH-SY5Y grades 1 to 4d according to Yamamoto (Cancer; 51:2175-80, cells but not in 590-nm irradiated cells, resulted in a decrease 1983). Of these 42 cases, 29 were grades 2+3 and 13 were of caspase-3 and -9 activities. Taken together, 590-nm grades 4c+4d. No obvious immunostaining of stathmin was irradiation can protect neuronal death through blocking observed in normal tongue epithelium. Positive expression NO-induced mitochondrial apoptotic pathway. Thus, specific of stathmin in tumor cells was detected in all 42 cases of wavelength of irradiation could be used for prevention of tongue cancer. However, the expression pattern was clearly neurodegenerative disorder. different depending on mode of invasion. In cases of grades 2+3, stathmin expression was limited to the peripheral cells 25. Glycyrol inhibits phorbol ester-induced NF-κB- of the cancer nests, with very little stathmin expression dependent transcriptional activity and induces observed in cancer cells located inside of the cancer nests. In gene expression to tumor suppression through p53- cases of grades 4c+4d, strong positive staining for stathmin dependent apoptosis: A view of gene expression was detected throughout the small cancer nests. Extensive extending from apoptosis into tumor suppression staining was observed in tumor cells invading diffusely into within HEK 293T cells the deepest portion of the tissue. We conclude that expression SungGa Lee, Won-Bong Lim, In-Ae Kim, Min-A of stathmin in human tongue cancer cells correlates with the Jung, Ok-Joon Kim, Hong-Ran Cho mode of invasion. Thus, stathmin may play an important Chonnam National University, Gwangju, Korea role in the invasion of tongue cancer. Glycyrrhiza uralensis (Leguminosae) has long been known as an anti-inflammatory agent for gastric ulcers, 24. Anti-apoptotic pathway of 590-nm irradiation arthritis, and rheumatism. The flavonoid glycyrol isolated in SNP induced SH-SY5Y neuroblastoma cell lines from Glycyrrhiza uralensis dramatically inhibits phorbol InAe Kim1, WonBong Lim1, SungGa Lee1, MinA ester (PMA)-induced NF-κB-dependent transcriptional Chung1, OkJoon Kim1, Gwon Tae Youn2, HongRan activity, as determined by luciferase activity in HEK 293T Choi1 cells. To investigate the global gene expression profiling Department of Oral Pathology1, 2nd stage of brain Korea 21 for modulated by glycyrol, we performed high-density School of Dentistry, Dental Science Research Institute, Chonnam oligonucleotide microarrays, followed by validation using National University, Gwangju, Korea and Chang Sung Health RT-PCR. Among the 33,096 genes that were screened in the center2, Jeollanam-Do, Korea microarray, 1,322 genes [?4% of the genome, flag value < The nitric oxide (NO) is a major factor contributing to 5000, signal noise (SN) > 3, and P value < 0.05] were found the loss of neurons in ischemic stroke, demyelinating to be differentially expressed between un-treated and PMA- diseases, and other neurodegenerative disorders. However, it treated HEK 293T cells. Among these candidates, 216 genes is known that NO is not functioning as a direct neurotoxin. [?0.65% of the genome, fold change (FC) of PMA to Control − NO combined with superoxide (O2 ) by the diffusion- > 3], which were highly up-regulated in cells treated with controlled reaction, formed a peroxinitrite anion (ONOO−), PMA, were significantly down-regulated in cells treated and this species has been shown to contribute to oxidative with both glycyrol and PMA. Our microarray analysis signaling and damage. ONOO− could trigger apoptosis in showed that glycyrol inhibited phorbol ester-induced many cell types, whether ONOO− acts directly as an oxidant NF-κB-dependent transcriptional activity in inflammatory- or stimulates the intracellular apoptotic signal. However, the related gene expression. RT-PCR analysis based on mechanism by which ONOO− induces apoptosis is unclear microarray data also showed that NF-κB-dependent genes although subsequent formation of reactive oxygen species such as CCL 2, CCL 7, CD 44, HSPB 8, and TIMP 1 in (ROS) has been suggested. The aim of present study is to addition to NF-κB itself were significantly down-regulated investigate the anti-apoptotic pathway by blocking of by glycyrol. Treatment with glycyrol also inhibited I-κB ONOO− production through scavenging of ROS with degradation induced by PMA, suggesting that glycyrol has a specific wavelength of light irradiation. To achieve this potential anti-inflammatory activity by modulating NF-κB- object, the anti-apoptotic effect of specific wavelength dependent pathways. Also, the microarray data showed that irradiation in Sodium Nitroprusside (SNP)-treated SH-SY5Y glycyrol-induced expression of genes related to tumor cells was investigated by MTT, DNA fragmentation, flow suppression and apoptosis induction including the p53 gene. cytometric assay, western blot assay, and caspase activity Overall, glycyrol appears to induce gene expression of assay. Also, NO, ROS, or ONOO− level was measured to tumor suppression genes by p53-dependent apoptosis in observe the changes of NO involved in radicals. Our results HEK 293T cells. Therefore, it is expected that glycyrol is a showed that the cell viability was reduced to around 60% of novel candidate as a tumor suppressor with the mechanism control by SNP treatment, and relieved of 80% by 590-nm of apoptosis induction. irradiation. The apoptotic cells were observed by flow 72 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology

26. Ameloblastic carcinoma of the jaw: Clinico­ , an or an ameloblastoma. pathologic study of 5 cases CT showed bucco-lingually undulating expansile lesion Byoung-Chan Jo, Seong-Doo Hong, Jae-Il Lee, Sam- with corticated margin from the left posterior mandibular Pyo Hong body to the anterior ramus, including #46, #47 and #48. The Department of Oral Pathology, School of Dentistry, Seoul mass containing in the lesion showed slightly lower National University, Seoul, Korea attenuation than muscle, leading to the impression of an Ameloblastic carcinoma (AC) is a malignant counterpart ameloblastoma. The mass specimen obtained from surgical of ameloblastoma, histologically characterized by ameloblas­ excision was composed of 3 sac-like structures, measuring tomatous feature with obvious cytologic atypia. We reviewed 4.1 cm, 1.8 cm and 1.4 cm in diameter, respectively. One sac 5 cases of AC clinicopathologically according to age, sex, was tightly attached to the #46, resembling a dentigerous location and clinical findings. They occurred at the age from cyst. Microscopic examination showed a large number of 46 to 75 years with an average age of 60. Four tumors cholesterol clefts in association with hemorrhage, hemo­ occurred in male and one in female, showing predilection for siderin pigments, and foreign body giant cells. There was no male. While four cases occurred in the maxilla, including evidence of a cyst or other lesions. CG should be taken into three in the molar area and one in the premolar area, only differential diagnosis in addition to odontogenic cysts and one case occurred in the . Three patients presented tumors when a radiographically well-circumscribed mul­ a painful large oral ulcer at the time of admission, and other tilocular radiolucent lesion occurs. 2 patients presented swollen mass around the gingivae and alveolar bone. The duration of symptoms before final 28. Effects on proliferation, migration and invasion diagnosis ranged from 4 months to 2 years. Radiographically, of CXCR-4 down-regulation in oral cancer cells they showed poorly demarcated radiolucent lesion with Ji-Soo Hong, Kyoung-Ok Hong, Jae-Il Lee, Sam-Pyo irregular bony destruction. Microscopically, lymph node Hong, Seong-Doo Hong metastasis was confirmed in one case, but not in others. The Department of Oral Pathology, School of Dentistry, Seoul tumors showed basic histologic feature of an ameloblastoma National University, Seoul, Korea with apparent cytologic atypia such as pleomorphism, Chemokines receptors are seven-transmembrane, G- hyperchromatism and atypical mitosis. Individual neoplastic coupled proteins. Recent data suggest that the interactions cells displayed wide variations from case to case, showing between chemokines and their receptors are also critical clear cells with faint PAS-positive granules, isolated round components in the regulation of tumor progression and cells with abundant cytoplasm, squamoid cells, and polygonal metastasis in breast cancer and other tumors. Above all, cells. Immunohistochemical survey was done in 2 cases, all chemokine receptor CXCR-4 and its ligand stromal cell- of which showed positivity to pan-cytokeratin antibody and derived factor-1 (SDF-1) have been shown to mediate organ p53 antibody, but negativity to HMB-45, S-100, and SMA. specific metastasis by creating a chemotatic gradient between One case recurred twice during 2 years after surgery, but 3 the primary tumor site and the metastatic site. CXCL12/ cases showed no evidence of recurrence and metastasis. It CXCR4 signal is implicated in tumor proliferation, survival, seems to us that ACs have the potential of metastasis and vascularization, metastasis, and immunosuppression. How­ recurrence. ever, there are few reports about the expression pattern of chemokine in oral cancer. In this study, we investigated the 27. Cholesterol granuloma in the mandible - case mRNA expression and protein level of CXCR-4 in several report oral cancer cell lines. Specially, KOSCC-25B and KB cell Ji-Hong Kim, Seong-Doo Hong, Jae-Il Lee, Sam-Pyo showed higher CXCR-4 expression, whereas the rest of the Hong cell lines rarely expressed. Also, we induced down-regulation Department of Oral Pathology, College of Dentistry, Seoul of CXCR-4 expression using small interfering RNA (siRNA) National University, Seoul, Korea in KOSCC-25B. Reduction in CXCR-4 expression by Cholesterol granuloma (CG) occurs frequently in siRNA decreased the proliferation and migration of KOSCC- association with chronic middle ear diseases, particularly 25B cells. These data suggest that CXCR-4 down-regulation diseases in the mastoid antrum and air cells of the temporal in oral cancer may be an effective tool for anti-cancer bone, and much less frequently in paranasal sinuses. It strategy. occurs frequently secondary to massive hemorrhage of oral and paraoral cysts. However, it has never been reported to 29. Downregulation of epithelial-mesenchymal occur solely without any association of preexisting lesion in tran­sition-related Snail1 and Twist, not Sip1 in oral the mandible. We experienced the development of an squamous cell carcinoma cells by inhibition of Akt unusual cholesterol granuloma in the mandible. A 70-year- activity old female patient presented a diffuse hard swelling on the Kyoung-Ok Hong, Ji-Soo Hong, Jae-Il Lee, Sam-Pyo left mandibular area with lymphadenopathy in the left Hong, Seong-Doo Hong cervical lymph node. Radiographic examination showed a Department of Oral Pathology, School of Dentistry, Seoul well-circumscribed multilocular radiolucency resembling National University, Seoul, Korea soap bubble appearance with tooth displacement and root Epithelial-mesenchymal transition (EMT) plays a pivotal resorption, leading to the radiographic impression of a role in the conversion of early stage tumors into invasive Oral Med Pathol 12 (2008) 73 malignancies, and such changes are associated with aberrant cytometry, laser scanning confocal microscope, and RT-PCR. expression of developmentally regulated and mesenchymal Results showed that 635-nm irradiation and existing COX cell-related genes. Among the mechanisms largely associated inhibitors inhibited expression of COX and PGE2 release. with the metastatic conversion of epithelial cells and the Unlike indomethacin and ibuprofen, 635-nm irradiation led EMT, one of the prominent changes is the loss of E-cadherin- to a decrease of ROS levels and mRNA expression of mediated cell adhesion. Indeed, most of the E-cadherin cytosolic phospholipase A2 (cPLA2) and secretary phos­ repressors, including Snail1, Twist, and Sip1, seem to fulfill pholipase A2 (sPLA2). Taken together, 635-nm irradiation, this requirement, and they induce complete EMT when unlike indomethacin and ibuprofen, can directly dissociate overexpressed in epithelial cell systems. The Akt/PKB the ROS. This inhibits cPLA2, sPLA2, and COX expression, family of kinases is a downstream effector of phosphati­ and results in the inhibition of PGE2 release. Thus, we dylinositol 3-kinase (PI3K) and is frequently activated in suggest that 635-nm irradiation inhibits PGE2 synthesis like human cancers including oral cancer. Recently, activation of COX inhibitor and appears to be used as an anti-inflammatory the PI3K/Akt axis is emerging as a central feature of EMT. tool. However, the specific interactions of the Akt signaling with other E-cadherin repressors like Twist and Sip1 during EMT 31. Hookah (water pipe) epidemic in the Indian in OSCC cells have not been clearly elucidated. In the sub­continent present study, we investigated whether Akt inhibition by PIA Simarpreet Sandhu1, Jagpreet Sandhu2, Simran Nijjar3 treatment, structurally modified phosphatidylinositol ether G.I.D.S.R.1, Ferozepur, India; D.I.R.D.S.2, Faridkot, India; and lipid analogues (PIAs), could downregulate the Snail1, Sip1 Amritsar3, India and Twist expression in oral cancer cells. We showed that The World Health Organization Study group on Tobacco inhibition of phosphorylated Akt by PIA could effectively Product Regulation (TobReg) has issued in 2005 an increase GSK3 phosphorylation and lead to downregulation “Advisory Note” entitled “Waterpipe Tobacco Smoking: of Snail and Twist expression. We also showed that Akt Health Effects, Research Needs and Recommended Actions inhibition reduced tumor cell migration and invasion. by Regulars”. This important report, the first ever prepared Interestingly, Akt inhibition did not affect ERK phos­ by WHO, has considered ‘waterpipe’ or ‘hookah’ smoking a phorylation and Sip1 expression. All of these findings global public health threat. The global tobacco epidemic suggest that Akt signaling may induce the EMT through may kill 10 million people annually in the next 20-30 years activation of Snail1 and Twist, not Sip1 in OSCC cells. with 70% of these deaths occurring in developing countries. Current research, treatment and policy efforts focus on 30. The anti-inflammatory mechanism of 635-nm cigarettes, while many people in developing regions (Asia, light-emitting-diode irradiation compared with Indian subcontinent, Eastern Mediterranean) smoke tobacco existing COX inhibitors using waterpipes. Waterpipes or hookah are increasing in Wonbong Lim1, SungGa Lee1, Inae Kim1, Mina popularity and more should be learnt about them so that we Chung1, Misook Kim1, Saewoo Han1, Hoisoon Lim1, can understand their effects on public health, curtail their Jinsoo Park2, Hongran Choi1, Okjoon Kim1 spread and help their users quit. Medline search using Department of Oral Pathology1, 2nd Stage of Brain Korea 21 keywords “Waterpipe”, “Hookah” and “Indian subcontinent” for School of Dentistry, Dental Science Research Institute, was conducted with special emphasis on chemistry of Chonnam National University, Gwangju, Korea and Biophoton2, smoke, health related effects, smoking patterns, history, Gwangju, Korea gender and under age use aspects, prevention, and research Inhibition of cyclooxygenase (COX) and prostaglandin needs. Every identified published study related to waterpipe E2 (PGE2) protects cells against cell injury in specific was included. We found that research regarding waterpipe pathophysiological situations: inflammation and oxidative epidemiology and health effects is limited. Hookah is stress. Although the anti-inflammatory effects have been increasing globally particularly in Indian subcontinent reported in clinical fields for specific wavelength irradiation where perceptions regarding health effects and traditional during wound healing, the physiological mechanism has not values may facilitate use among women and children. We been clarified yet. The aim of the present study wasto conclude that more scientific documentation and careful investigate the anti-inflammatory mechanism of 635-nm analysis is needed before the spread of hookah use, its health light-emitting-diode (LED) irradiation compared with effects can be understood, and empirically guided treatment existing COX inhibitors. The present study investigated and public policy strategies can be implemented. anti-inflammatory effects of 635-nm irradiation on PGE2 release, COX and phospholipase A2 (PLA2) expression, and 32. Recurrent peripheral ameloblastic carcinoma reactive oxygen species (ROS) dissociation in arachidonic with lung metastases - case report acid (AA)-treated human gingival fibroblast (hGF). These Shin-Yu Lu1, Shun-Chen Huang2, Cheng-Hua Huang3 results were compared with their existing COX inhibitors: Department of Family Dentistry1, Department of Pathology2, 3 indomethacin and ibuprofen. The PGE2 release was measured and Department of Oncology , Chang Gung Memorial Hospital, by enzyme immunoassay, the COX expression was measured Kaohsiung Medical Center, Chang Gung University, College of by western blot and reverse transcriptase-polymerase chain Medicine, Kaohsiung, Taiwan reaction (RT-PCR). ROS level was measured by flow Peripheral ameloblastic carcinoma (PAC) is a rare 74 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology malignant neoplasm derived from either extraosseous galectin-1 in epithelia was positively correlated with the remnants of the dental lamina or basal cells of the oral precancerous lesion progression from lower-risk prema­ epithelium. PAC is defined as a type of extraosseous lignant lesion (0%), higher-risk premalignant lesion (16%), ameloblastoma with histologic evidence of malignant to OSCC (35%) (P = 0.03). By double immunofluorescent transformation of the epithelial component in both primary staining, we found that galectin-1 immunoreactivities are and metastatic lesions. PAC has the potential to invade mostly co-localized with the vimentin (the marker of the jaw bone and metastasize to regional lymph nodes or fibroblast). In conclusion, our findings suggest that galectin-1 distant organs. This article reported an unusual case of PAC up-regulation may be a predictor especially in early presenting initially as a polyp on the buccal gingiva of the progression of oral cancer. right lower retromolar area of a 56-year-old edentulous male patient. The lesion was excised, and a PAC was 34. Tuberculosis of the head and neck: a review of diagnosed. Because the wound healing was uneventful, the 18 cases patient refused further wide excision. Three years later, the Wen-Chen Wang1, Yuk-Kwan Chen1,2, Jin-Yi Chen1, lesion recurred locally without underlying bone invasion Cheng-Chung Lin1, Li-Min Lin1,2 but with multiple metastases to the lungs, bronchus, and Department of Oral Pathology and Diagnosis1, Kaohsiung pleura. Histologically, primary, recurrent, and metastatic Medical University Hospital and Department of Oral Pathology2, tumors excised or biopsied showed typical features of Kaohsiung Medical University, Kaohsiung, Taiwan PAC. Chemotherapy with cisplatin and epirubicin was Tuberculosis (TB) is a worldwide health problem, but given to the patient to control the recurrent and metastatic TB of the head and neck could be a diagnostic challenge. tumors, and it led to marked regression of the recurrent oral The aim of the present study was to review the clinical tumor and bilateral pulmonary carcinomatosis. However, features of TB of the head and neck. The information chemotherapy was withheld because of anthracycline- acquired may aid the earlier clinical diagnosis. A total of 12 related cardio­myopathy. The patient rapidly deteriorated male and 6 female patients with TB of the head and neck and died of respiratory failure due to pneumonia and tumor area diagnosed by surgical biopsy were collected between progression 17 months after the diagnosis of recurrent PAC 1991 and 2007 in the Department of Oral Pathology and with lung metastasis. We suggest that wide surgical excision Diagnosis of Kaohsiung Medical University Hospital. of the primary tumor is needed after a pathological diagnosis Medical charts were carefully reviewed. We found that of PAC, and a patient with PAC requires a meticulous among these 18 patients, 11 (61.1%) were above the 5th follow-up. Because the undifferentiated tumor cells of decade and 3 (16.7%) were in the 1st decade. 13 patients had PAC are sensitive to the two chemotherapeutic agents we oral lesions, and 2 had multiple sites of occurrence. The gave to this patient, combination chemotherapy may be the most common locations of the oral lesions were vestibule of treatment of choice for recurrent and widespread PAC. the mandible (n = 3) and buccal mucosa (n = 3), followed by edentulous ridge of the mandible (n = 2), lower (n = 2), 33. Galectin-1 up-regulation is associated with pre­ retromolar area (n = 1), tongue (n = 1), soft (n = 1), cancerous transformation during oral carcinogenesis and tuberosity (n = 1). The most common clinical mani­ Yuh-Ling Chen, Fu-Ching Ciou, Ying-Tai Jin, Tung- festation was delayed healing of oral ulcers (69.2%), and Yiu Wong usually clinically misdiagnosed as oral squamous cell Institute of Oral Medicine, National Cheng Kung University, carcinoma. On the other hand, 5 patients had cervical Tainan, Taiwan lesions. Significantly, there were three patients aged 6, 8 and Oral squamous cell carcinoma (OSCC) is the most 17 years, respectively. Further, two patients were coexistent common cancer in the head and neck region, and its with metastatic squamous cell carcinoma in cervical lymph transformation is generally from normal epithelia to nodes. Four patients with active pulmonary lesions and 1 precancerous lesions (PML), and then some of these lesions patient with old TB were identified from the 12 chest may change into OSCC. Among these precancerous radiographs. We found that TB of the head and neck seems transformations, verrucous hyperplasia and severe dysplasia, not to be a rare lesion. Due to the risk of infection and and carcinoma in situ are thought to be more intriguing and transmission of TB to dental care providers, we emphasize dangerous. Altered expression of galectin-1 in oral cancerous the importance of identification of the head and neck lesions, tissue was detected (Chiang et al., 2007) but their roles in the especially for the delayed healing wounds and neck lumps. progression from precancerous to cancerous stages remained unclear. In this study, we analyzed the immunohistochemical 35. An alternative iron oxide nanoparticles drug (IHC) profiles of galectin-1 in human oral verrucous carrier for oral cancer therapy hyperplasia, severe dysplasia, and carcinoma in situ. For Ping-Chin Wu1, Tsung-Lin Tsai1, Fong-Yu Cheng2, further examination of cellular distribution of galectin-1 in Chen-Sheng Yeh2, Dar-Bin Shieh3 the precancerous tissue, immunofluorescent assay was Institute of Basic Medical Sciences1, Department of Chemistry2, performed. IHC staining showed that galectin-1 was highly Institute of Oral Medicine and Molecular Medicine3, National expressed in 16% of epithelial cells and 56% in stromal layer Cheng Kung University, Tainan, Taiwan of the precancerous cases, and the overexpression rate was Cancers remain as the major course of human death in increased in the cancer group. The overexpression of many countries around the world. Different strategies have Oral Med Pathol 12 (2008) 75 been proposed to eliminate cancer cells and to avoid 37. Ameloblastoma with an unusual radiographic potential collateral damages to healthy tissues. Nanoparticles fea­ture - case report have recently been extensively investigated for their potential Yi-Juai Chen, Yu-Feng Huang biomedical applications in both diagnostics and therapeutics School of Dentistry, Chung Shan Medical University and due to their distinct physical and chemical properties Department of Dentistry, Chung Shan Medical University compared to the bulk status. In this study, an aqueous phase Hospital, Taichung, Taiwan synthesis of superparamagnetic Fe3O4 nanoparticles was Ameloblastoma is the most common benign odontogenic applied for anticancer therapy. They have a typical diameter tumor. The majority of cases occur in the posterior of 6.2 ± 1.1 nm. Zeta potential analysis revealed their mandible. Radiographically, ameloblastomas are usually positive charge surface. Chemical modification of the well-defined radiolucency with or without a sclerotic surface was performed in the particle surface to enable border. Common to all subtypes of ameloblastoma is the affinity adsorption of anticancer drug - Epirubicine. Cancer palisading cells mimicking that of ameloblasts of the cytotoxic efficacy was then evaluated by WST-1 assay in enamel organ. Another typical feature is the budding of various oral cancer cells. In conclusion, this study demon­ tumors from neoplastic foci in a pattern reminiscent of tooth strated an alternative strategy for loading and releasing development. However, no hard tissue formation is present. anticancer drugs for potential combination of target therapy Although histologically benign, the tumor has a reputation using a porous iron oxide nanoparticle as the core carrier. of high recurrent rate because of its local aggressiveness. Thus, wider surgical excision is the treatment of choice. 36. Areca nut extracts increase expression of Here, we reported a case of ameloblastoma with an unusual proinflam­matory cytokines, TNF-α and IL-1β, by radiographic pattern. A 34-year-old woman who visited immune cells our dental clinic complained of a painless swelling in the Lien-Yu Chang, Hsiao-Ching Wan, Shan-Ling Hung right mandible for more than five months. At examination, National Yang-Ming University, Taipei, Taiwan a bony hard, asymptomatic swelling was noted at the #43 Areca quid chewing is a very popular habit in Taiwan and #44 area, mainly on the buccal gingiva. Radiographic and is associated with some oral diseases. Inflammation examination revealed a well-defined radiolucent-radiopaque orchestrates the local environment, is a crucial event for lesion involving #43 and #44. All involved teeth were disease progression. Inflammatory cytokines, IL-1b and vital. A clinical impression of benign fibro-osseous lesion TNF-a are often present simultaneously in chronic was obtained, including ossifying fibroma. Since patient inflammatory diseases and known to underpin tissue damage. refused an incisional biopsy, a total tumor enucleation was The purpose of this study was to investigate the role of areca performed with trimming of surrounding bone. Histological in production of cytokines, TNF-α and IL-1β, by areca nut examination showed features of typical ameloblastoma in extracts (ANE)-stimulated immune cells (peripheral blood most areas. However, spherical calcifications were observed mononuclear cells, PBMCs). PBMCs were isolated from in focal area associated with more acanthomatous feature venous blood of healthy donors with informed consent. of the lesion. Thus, a final diagnosis of ameloblastoma was Expression of TNF-α and IL-1β by treated immune cells rendered. Healing was uneventful. However, follow-up was examined using enzyme-linked immunosorbent assay radiographs showed rapid recurrence of the lesion within and reverse transcription-polymerase chain reaction. The 5 months. Since there is still no clinical sign of recurrence, results demonstrated that either ripe ANE (rANE) or tender close follow-up is still on-going. ANE (tANE) significantly enhanced the production of IL-1b and TNF-a in cultured cells by a dose-dependent manner. At 38. Hemodialysis-related lingual amyloidosis: report the same concentrations used, rANE induced more cytokine of two cases production than tANE. The mRNA expression was also Jehn-Shyun Huang, Ying-Tai Jin, Tong-Yui Wong, increased by either rANE or tANE. The increased secretion Ken-Chung Chen, Shih-Ming Wong of TNF-α and IL-1β by treated cells could be attenuated by National Cheng Kung University, Tainan, Taiwan a higher concentration (50 μM) of antioxidants, PDTC or Hemodialysis-related amyloidosis is a serious curcumin. However, only the production of IL-1β by tANE- complication of long-term dialysis treatment due to the treated cells could be decreased significantly by 10 μM of deposition of beta-2-microglobulin, predominantly in curcumin. We conclude that the increased expression of key osteoarticular structures. It is rarely found in visceral organs inflammatory mediators, TNF-α and IL-1β, by areca-treated or the tongue. When such extra-articular involvement immune cells may play an important role in creating a happens, it denotes a more advanced stage of the disease. In sustained chronic inflammatory oral condition in areca quid this report, we presented two cases of lingual amyloidosis chewers, hence promote oral disease progression. Moreover, who received hemodialysis for more than 10 years. The induction of oxidative stress is a critical event in this areca- diagnosis led to the recognition of the disease process of associated immune deterioration. beta-2-microglobulin deposition in these patients. One patient died and the other survived 2 years after the diagnosis. The clinical appearance of the lingual amyloidosis was one with nodular type and the other with diffused type combined with . Both cases received biopsy for 76 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology pathological prove and the microscopic pictures revealed small primary tumors. Galectin-1 is one significantly amorphous amyloid deposition in the submucosal layer. upregulated tumor-associated protein in many neoplasms. Occasionally, lingual amyloidosis may appear to the oral To determine the clinical significance of galectin-1, we and maxillofacial pathologists as their first clinical clue of analyzed its expression in clinical samples by reverse such systemic disease process. The clinicians might meet transcription-quantitative polymerase chain reaction assay diagnostic challenge due to its rarity and the unfamiliar to analyze microdissected tissue and immunohistochemical picture of the disease process. The cardinal signs and assay. Galectin-1 was significantly upregulated in stromal symptoms may promptly arouse of the suspicious of such parts during early-stage OSCC and in epithelial parts at disease process. Making an appropriate diagnosis might halt the metastatic stage in both assays. Survival analysis and a the progression of the disease to the more severe stage. Cox’s proportional hazards model showed that synchronous upregulation of galectin-1 protein and mRNA was correlated 39. Nevoid basal cell carcinoma syndrome - case with worse disease-free survival in early-stage OSCC (P = report 0.024 and P = 0.047, respectively). Our findings suggest that Yi-Juai Chen, Yi-Tzu Chen, Bour-Chang Wu, Chih-Yu galectin-1 upregulation at the tumor invasion front may be a Peng, Ming-Yong Chou, Yu-Feng Huang predictor of early metastasis in oral carcinogenesis and laser Department of Oral and Maxillofacial Surgery, College of Oral microdissection is a useful tool in clinical analysis of target Medicine, Chung Shan Medical University, Taichung, Taiwan gene in either epithelial or stromal component. Nevoid basal cell carcinoma syndrome is an autosomal dominant inherited disease. The prevalence of the syndrome 41. Expression of inhibitors of apoptosis family in is estimated to be about 1 in 60,000. One of the major DMBA-induced hamster buccal-pouch squamous- manifestations of these patients is multiple odontogenic cell carcinogenesis is associated with p53 accumu­ keratocysts of the jaws, in addition to multiple nevus-like lation and epigenetic mechanism pigmentations which represent basal cell carcinomas. Other Shue-Sang Hsue, Yuk-Kwan Chen, Li-Min Lin commonly seen clinical features include calcification of falx Department of Oral Pathology, School of Dentistry, College of cerebri, bifid rib, and palmar/plantar pits. Many mutations in Dental Medicine, Kaohsiung Medical University, Kaohsiung, PTCH gene have been reported. Here we presented a case of Taiwan and Kaohsiung Medical University Hospital, Kaohsiung, Gorlin syndrome in a 15-year-old girl who initially presented Taiwan with a well-defined radiolucent lesion associated with an Apoptosis is regulated by a number of inhibitory or impacted #33 in July, 2006. During the course of examination, stimulatory factors. One such family of anti-apoptotic bifid of right 4th rib and calcified falx cerebri were noted. proteins is inhibitors of apoptosis (IAP) family. Eight human The jawbone lesion was enucleated and the histopathologic IAP family members have been recognized so far, including report was an with an impaction of neuronal apoptosis inhibitory protein (NAIP), X-linked #33. Three months later, additional well-defined radiolucent inhibitors of apoptosis protein (XIAP), cellular inhibitors of lesions were seen on panoramic radiograph associated with apoptosis protein 1 (cIAP1), cellular inhibitors of apoptosis developing #38 and #48. Both lesions were again removed protein 2 (cIAP2), survivin, baculoviral IAP repeat-con­ and histopathologic reports were also odontogenic kera­ taining ubiquitin-conjugating enzyme (BRUCE) (apollon), tocysts, thus confirming the diagnosis of nevoid basal cell livin (ML-IAP, KIAP), and IAP-like protein 2 (ILP-2). The carcinoma syndrome. Initial genetic search on mutations of relationship between the expression of IAP family and p53 PTCH showed 2 mutations. One was on exon 12 (c.1686 bp status as well as epigenetic alterations in chemically-induced CRT) and the other on intron 13 (g.91625bp CRG). No basal experimental oral carcinogenesis is not completely under­ cell carcinoma was discovered on the skin, nor was stood. Therefore, the objective of the present study was to recurrence of odontogenic keratocyst identified. Patient was investigate the expression of five IAP members (survivin, currently under close follow-up. XIAP, cIAP1, cIAP2 and NAIP) and their relationship with p53 status and epigenetic mechanism in 7,12-dimethylbenz[a] 40. Galectin-1 mRNA levels quantified by reverse anthracene (DMBA)-induced hamster buccal-pouch squamous- transcriptase-polymerase chain reaction in micro­ cell carcinogenesis. Fifty outbred, young (6 weeks) male dissected oral carcinoma tissues are correlated with Syrian golden hamsters were randomly divided into three histological grade of both epithelial and stromal experimental groups (10 animals each) and two control components groups (each with 10 animals). The pouches of the three Wei-Fan Chiang1, Yuh-Ling Chen2, Shyun-Yeu Liu1, experimental groups were painted bilaterally with a 0.5% Ying-Tai Jin2 DMBA solution three times a week for 3, 7 and 14 weeks. Chi-Mei Medical Center1, Tainan, Taiwan and National Cheng The treatment protocol for animals in one of the control Kung University Hospital2, Tainan, Taiwan groups was identical with only mineral oil applied while Metastasis is a final and fatal step in the progression another control group remained untreated throughout the of oral squamous cell carcinoma (OSCC) and usually experiment. Immunohistochemical staining, semi-quan­ accompanies a large primary tumor. Metastasis may also titative RT-PCR and PCR-based methylation assay were come from a small primary tumor and progress rapidly; performed. Neither survivin nor cIAP2 (protein and mRNA) therefore, early diagnosis is important for patients with could be detected in any of the control tissues whereas both Oral Med Pathol 12 (2008) 77 mRNA and protein of survivin and cIAP2 were significantly (ANA), anti-smooth muscle (SMA), anti-mitochondrial increased for 3-, 7- and 14-wk DMBA-treated pouches (AMA), anti-gastric parietal cell (GPCA), anti-thyroglobulin compared with the control pouches. mRNA and protein (TGA), and anti-thyroid microsomal antibodies (TMA) in expression of XIAP, cIAP1 and NAIP were noted for both small groups of patients with oral lichen planus (OLP). In the control and 3-, 7- and 14-wk DMBA-treated pouches. this study, we used an indirect immunofluorescence technique However, both mRNA and protein expression of XIAP, to determine the serum levels of ANA, SMA, AMA and cIAP1 and NAIP for the 3-, 7- and 14-wk DMBA-treated GPCA, and a semi-quantitative microtiter particle aggluti­ pouches were elevated significantly as compared with the nation test to assess the serum levels of TGA and TMA in a control pouches. Moreover, the mRNA expression of these large group of 278 (46 men and 232 women) Chinese OLP five IAP members was significantly correlated to each other. patients and in 53 healthy control subjects. We found that p53 (both protein and mRNA) was not detected in any serum autoantibodies were present in 172 (61.9%) of the 278 control tissues whereas p53 expression for 3-, 7- and 14-wk OLP patients: 89 (32.0%) had only one autoantibody, 42 DMBA-treated pouches was significantly higher than that of (15.1%) two autoantibodies, 26 (9.4%) three autoantibodies, the control tissues. The mRNA expression of p53 was 14 (5.0%) four autoantibodies, and 1 (0.4%) five autoanti­ significantly correlated with that of the five IAP members. bodies. The frequencies of presence of serum ANA (27.0%, Direct sequencing revealed a point mutation (C→G) of p53 P = 0.001), SMA (9.7%, P = 0.036), GPCA (27.7%, P = for pouch tissues treated with DMBA for 3 wks and 7 wks as 0.000), TGA (21.6%, P = 0.001) and TMA (24.5%, P = compared with the control tissues. There was a wide 0.000) in OLP patients were significantly higher than those variation of the p53 sequence of the 14-wk DMBA-treated (5.7%, 0%, 1.9%, 1.9% and 1.9%, respectively) in healthy pouch tissues compared with the control tissues. Furthermore, control subjects. Furthermore, incidences of approximately all the control tissues had a survivin- and cIAP2-methylated 34-48% overlap of organ-specific and non-organ-specific allele, whereas all the DMBA-treated tissues showed no serum autoantibodies were found in our OLP patients with evidence of survivin- and cIAP2-methylation. On the other one or more organ-specific or non-organ-specific serum hand, both the control and DMBA-treated pouches showed autoantibodies. In conclusion, present and previous findings no evidence of XIAP-, cIAP1- and NAIP-methylation. In of a marked female predilection, the presence of multiple conclusion, our results suggest that IAP family could play an serum autoantibodies and hypergammaglobulinemia, and an important role in DMBA-induced hamster buccal-pouch overlap of organ-specific and non-organ-specific autoanti­ squamous-cell carcinogenesis and that their expression may bodies in OLP patients suggest that OLP may be an be modulated by both genetic (mutant p53) and epigenetic autoimmune disease. mechanisms. 44. Peripheral ossifying fibroma: a clinicopathologic 42. Carcinosarcoma (malignant mixed tumor) of study of 27 cases the parotid gland: A case report and review of the Hsien-Yen Hung, Chia-Chuan Chang, Julia Yu-Fong literature Chang, Chuan-Hang Yu, Yi-Ping Wang, Bu-Yuan Liu, Huan-Ching Su, Long-Chang Chang, Shin Nieh Chun-Pin Chiang National Defense Medical Center, Taipei, Taiwan Graduate Institute of Clinical Dentistry, School of Dentistry, and Carcinosarcoma is a biphasic neoplasm composed of Dental Department of National Taiwan University Hospital, both malignant epithelial and mesenchymal components. It College of Medicine, National Taiwan University, Taipei, Taiwan was first described in 1951 by Kirklin et al. This uncommon Peripheral ossifying fibromas (POFs) present as a neoplasm constitutes 0.04% to 0.16% of all salivary gland tumor-like growth on the gingiva or alveolar mucosa. They neoplasms and 0.4% of all malignant salivary gland are often misdiagnosed as a fibrous epulis, a pyogenic neoplasms. More than 60 cases of carcinosarcoma have been granuloma or a peripheral giant cell granuloma. Twenty- reported in English-language literature up to date. Although seven patients each with a POF diagnosed and treated in our there is evidence that some carcinosarcomas arise de novo, institute during the period from 1988 to 2006 were analyzed others appear to be formed through malignant transformation in this study. There were 16 female and 11 male patients. The from a previously benign pleomorphic adenoma. In this mean age of the 27 patients at the time of diagnosis was 38 report, we presented the histologic, cytohistologic, and years. The size of 27 POFs ranged from 5 to 30 mm in immunohistochemical findings of a carcinosarcoma of the greatest dimension with an average size of 15 mm. Clinically, salivary gland in a 57-year-old man. they usually presented as reddish, firm, pedunculated or sessile masses on the gingiva or alveolar mucosa. The 43. High frequencies and overlap of serum autoanti­ overlying mucosa was either completely intact (13 cases) or bodies in Chinese patients with oral lichen planus focally ulcerated (14 cases). The more common site for POF Andy Sun, Julia Yu-Fong Chang, Chun-Pin Chiang lesions was the premolar and molar region of the maxilla School of Dentistry and Dental Department of National Taiwan (52%). All patients finally received total surgical excision of University Hospital, College of Medicine, National Taiwan the lesion. Of the 27 POFs, 4 (15%) had single recurrence. University, Taipei, Taiwan Histologically, POFs showed trabeculae or islands of Previous studies have shown the presence of serum mineralized products in a cellular fibrous connective tissue autoantibodies including anti-epithelial cell, anti-nuclear stroma. The mineralized products were defined as woven 78 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology bone (22 cases), lamellar bone (6 cases), cementoid (12 46. Central ossifying fibroma: a clinicopathological cases), or amorphous material of dystrophic calcification (4 study of 28 cases cases). We conclude that POFs occur more commonly in Chia-Chuan Chang, Hsien-Yen Hung, Julia Yu-Fong female patients under 50 years old. The more common site Chang, Chuan-Hang Yu, Yi-Ping Wang, Bu-Yuan Liu, for POFs was the premolar and molar region of the maxilla Chun-Pin Chiang in this series. The recurrence rate for our POF lesions was Graduate Institute of Clinical Dentistry, School of Dentistry, and 15%. Dental Department of National Taiwan University Hospital, College of Medicine, National Taiwan University, Taipei, Taiwan 45. Metastatic prostate cancer in the mandibular Central ossifying fibroma (COF) is the most common trig­one region - case report benign fibro-osseous lesion of the jaw. This retrospective Hsien-Yen Hung, Hsin-Ming Chen, Chia-Chuan study evaluated the clinical and histopathological features of Chang, Julia Yu-Fong Chang, Chuan-Hang Yu, Yi-Ping 28 COFs in Taiwanese patients. Twenty-eight consecutive Wang, Bu-Yuan Liu, Chun-Pin Chiang cases of COF were collected from 1988 to 2006. Clinical Graduate Institute of Clinical Dentistry, School of Dentistry, and data and microscopic features of these cases were reviewed Dental Department of National Taiwan University Hospital, and analyzed. The mean age of patients at the time of College of Medicine, National Taiwan University, Taipei, Taiwan diagnosis was 34 years. There were 6 male and 22 female Oral metastatic tumors account for approximately 1% of patients. Twenty-six (93%) cases were found in the mandible oral malignant tumors and those to the jawbones are far and 2 (7%) in the maxilla. The most common site for COFs more common than those to the oral mucosa. In this report, were the molar region (17 cases, 61%), followed by the we presented the clinical course and histological feature of a premolar region (8 cases, 28%), and incisor/ canine region metastatic prostate adenocarcinoma to the mandibular (3 cases, 11%). Bone swelling or expansion (96%, 26/27) trigone region in a 72-year-old male patient. The patient was was the most frequent clinical presentation. Six (21%) COFs initially found to have a prostate adenocarcinoma in March presented as a radiolucent lesion, 17 (61%) as a mixed 2004. At that time, whole body bone scan discovered lesion, and 5 (18%) as a radiopaque lesion. No recurrence of multiple bone metastases to the left mandible, right first rib, the lesion was found after surgical excision in this series. upper T spine, and L3. Computed tomography of the pelvic Microscopically, COFs showed trabeculae of woven bone area revealed local tumor invasion and multiple lymphoade­ (25 cases) and/or lamellar bone (5 cases) and/or spherules of nopathies. Therefore, the prostate cancer was classified as cementoid (19 cases) in a cellular fibrous connective tissue being in stage IV and the patient had been treated by stroma. The stromal component was highly cellular in 21 palliative transurethral resection of the prostate and hormonal cases, moderately cellular in 7 cases, prominently vascular therapy with Leuplin and anti-androgen agents at the in 11 cases, and collagenous in 6 cases. COFs occur more Urology Department since March 2004. Two years later, a frequently in female patients and in patients in the second to firm, erythematous mass developed at the socket of #38 after fourth decade of life. The more commonly affected site is the tooth extraction. In addition, there was a high prostate- mandible, especially the molar region. The majority of COF specific antigen (PSA) level in patient’s blood. Biopsy of the lesions present as a well-defined, mixed lesion radio­ oral tumor showed solid nests of clear cells with prominent graphically. Most COFs can be treated by conservative nucleoli infiltrating in the gingiva and bone. Immunostaining surgical excision without subsequent recurrence. revealed that the tumor cells were strongly positive for PSA, and thus a metastatic prostate adenocarcinoma was diag­ 47. Idiopathic calcinosis over floor of the mouth and nosed. In May 2006, the patient was treated with radiotherapy submental region for control of oral tumor growth and cancer-caused bone Hsiao-Ting Huang1, Meng-Ling Chiang1, Chia-Jung pain. Abdominal and pelvic CT in July 2006 revealed Lin2, Yang-Ming Chang3 progression of cancer status. Due to poor control of the Department of General Dentistry1, Chang Gung Memorial cancer, he was further treated by systemic chemotherapy Hospital, Chang Gung University College of Medicine, Taipei, with Taxotere and Cisplatin. After chemotherapy, the oral Taiwan; National Taiwan University2, Taipei, Taiwan; and metastatic tumor regressed and the patient’s pain of the left Department of Oral and Maxillofacial Surgery3, Chang Gung mandible relieved. The patient was still under a regular Memorial Hospital, Chang Gung University College of Medi­ follow-up at both the Urology and Dental Departments cine, Taipei, Taiwan recently. Calcinosis is a term used to describe a group of disorders in which calcium deposits form. However, involvement of the oral cavity is rare. We reported a case of idiopathic calcinosis over floor of the mouth and submental region. A 35-year-old female came to our clinic with the complaint of a swelling over floor of the mouth. She stated that painful sensation was off and on and the mass progressively enlarged. Two granules were palpable both intraorally and extraorally. At first, the clinical diagnosis was and she was arranged for stone removal. However, the Oral Med Pathol 12 (2008) 79 stone-like masses were found two aside to sublingual gland expression was statistically significantly elevated in the and four under mylohyoid muscle. Sialogram revealed no malignant tissues compared with their corresponding salivary gland pathosis. Computer tomography showed premalignant leions. These results indicate that patients with multiple calcification masses at floor of the mouth. After pre-malignant oral lesions need long-term follow-up and stone removal and microscopic examination of specimens, may be monitored with IAP expression. the mass appeared to be calcification with focal ossification. The patient had been experienced surgeries for 3 times in the 49. Construction of single-domain antibody- past 15 years. She denied any systemic disease. Hence, the nanoparticle complex library system for MRI pathological diagnosis was idiopathic calcinosis. The imaging of cancers calcified masses may be recurrence or residual stones of Yi-Ling Chen1, Jianbing Zhang2, Chen-Sheng Yeh1, previous surgery. Long-term and regular follow-up will be Fong-Yu Cheng1, Wen-Yuan Hsieh3, Dar-Bin Shieh1 needed for this patient. National Cheng Kung University1, Tainan, Taiwan; Institute for Biological Sciences National Research Council of Canada2, 48. Malignant transformation in 1458 patients with Ottawa, Canada; and Material and Chemical Research Laboratories potentially malignant oral mucosal disorders: a Industrial Technology Research Institute3, Hsinchu, Taiwan follow-up study and its implication with inhibitors The purpose of this study was to construct an antibody- of apoptotic proteins nanoparticle complex library for in vitro magnetic resonance Yuk-Kwan Chen, Shui-Sang Hsue, Li-Min Lin imaging (MRI) and therapy for oral squamous cell carcinoma Department of Oral Pathology, Kaohsiung Medical University, (OSCC). Single domain antibody (SdAb) was constructed Kaohsiung, Taiwan into M13 phage library for screening oral cell carcinoma for It is generally accepted that the development of cancer in highly specific markers in collaboration with NRC of the oral mucosa is preceded by an identifiable non-invasive Canada. The genetic engineered 6-his tag in the C-terminal precursor lesion. Apoptosis is a complex and highly end of the SdAb enabled their self-assembly with iron oxide regulated process important for proper development and nanoparticles or Gd-dendrimer particles with Ni-NTA functioning of diverse roles such as tumorigenesis in surface modifications. The SdAbs were humanized to escape multicellular organisms and is executed by a family of adverse immunological response in the future human subject cysteine proteases known as caspase. The inhibitors of study. Thus screened M13 with high targeting efficiency apoptosis (IAP) are a family of proteins functioning as after bio-panning could be rapidly identified, amplified and intrinsic negative regulators of the aforementioned caspase assembled to form magic bullet for combined cancer cascade. The aim of this follow-up study was to estimate the diagnosis and therapy. In this study, SdAb recognized a rate and the time to transformation in a group of patients surface antigen that most OSCC lines were positive in from southern Taiwan with potentially malignant oral primary screening. The SdAb with a size only about one epithelial lesions and its implication with IAPs. The tenth of a normal IgG was found to target EGFR-2. The 3+ follow-up time is defined as the duration between the onset SdAb conjugated to Fe3O4 and Gd loaded dendrimer of the initial diagnosis and the occurrence of confirmed oral successfully produced image contrast in a dosage dependent cancer. A total of 1458 patients with histological diagnoses manner in MRI T1 and T2 sequences, respectively. of various premalignant oral lesions were followed up Histochemical and immunohistochemical staining of the between 1991 and 2001. The average age at initial diagnosis control normal and the cancer cell lines revealed specific was 47.5 years. The histological diagnoses were divided into targeting to the cancer cells. In summary, the combination of six categories: epithelial dysplasia with hyperkeratosis/ SdAb library and nanoparticles provide a new platform for epithelial hyperplasia (8.85%); epithelial dysplasia with clinical targeting diagnosis and therapy. The porous structure submucous fibrosis (2.54%); submucous fibrosis (27.57%); of the dendrimer further enabled drug or RNAi loading and hyperkeratosis/epithelial hyperplasia (29.01%); lichen delivery to the desired anatomic locale. The iron oxide could planus (9.80%), and verrucous hyperplasia (22.22%). The be engineered to produce heat at local regional area and thus best characterized IAP members are XIAP, NAIP, cIAP1, could be extended beyond diagnosis. cIAP2, and survivin. Their immunohistochemical expression in these 44 human premalignant oral lesions and their 50. Differential gene expression signature between corresponding malignant transformation lesions was primary and metastatic head and neck squamous investigated. Within the cohort of 1458 patients, 44 patients cell carcinoma progressed to oral cancer in the same site as the initial Chung-Ji Liu1-3, Kuo-Wei Chang1, Lan-Tin Kuo1, Hui- lesions with an overall transformation rate of 3.02% and a Wen Cheng4, Hsiao-Ching Chang4, Shu-Chun Lin1* mean follow-up time of 42.64 months. Eight of the 166 School of Dentistry1, National Yang-Ming University, Taipei, patients with dysplastic lesions and 15 of the 423 patients Taiwan; Department of Oral and Maxillofacial surgery2, Taipei with hyperkeratosis/epithelial hyperplasia progressed to MacKay Memorial Hospital, Taipei, Taiwan; MacKay Medicine, malignancy. The other patients with malignant transformation Nursing and Management College3, Taipei, Taiwan; and originated from various pre-cancerous oral lesions and Department of Medical Research4, Taipei MacKay Memorial conditions (submucous fibrosis, 8 of 402; lichen planus, 3 of Hospital, Taipei, Taiwan 143; verrucous hyperplasia, 10 of 324). Also, the IAPs Head and neck squamous cell carcinoma (HNSCC) is a 80 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology world-wide malignancy. This study aimed to identify the the combined increase in the RDW and MCV in patients attendant differential gene expression occurs as the disease with a sore mouth is still a useful indicator of possible progresses from cancer formation to metastatic HNSCC. vitamin B12 deficiency after folate deficiency and chronic Microdissection retrieved pure epithelial cells from paired diseases have been ruled out. primary tumors and cervical lymph node metastasis. cDNA microarray analysis and algorithm grouping identified 52. Insulin-like growth factor binding protein-5 differential mRNA expression of 301 genes. Quantitative (IGFBP-5) suppresses the tumorigenesis of oral reverse transcription-polymerase chain reaction analysis squamous cell carcinoma clarified the up-regulation of CCL19, CR2, EGR2, FUCA1, Pei-Shih Hung1, Shou-Yen Kao1,2, Chung-Ji Liu1,3, RGS1 and SELL as well as the down-regulation of IGFBP-6 Yin-Hua Shih1, Shuo-Tien Liu1, Shu-Chun Lin1* and KLK8 in nodal metastasis compared to primary tumors. Institute of Oral Biology1, School of Dentistry, National Yang- Immunohistochemistry confirmed the up-regulation of SELL Ming University, Taipei, Taiwan; Department of Dentistry2, and down-regulation of IGFBP6 in nodal metastasis relative Taipei Veterans General Hospital, Taipei, Taiwan; and to primary tumors. Interestingly, primary tumors exhibiting Department of Oral and Maxillofacial Surgery3, Taipei Mackay higher FUCA1 and SELL expression were associated with Memorial Hospital, Taipei, Taiwan significantly worse patient survival. In OECM-1 HNSCC Oral squamous cell carcinoma (OSCC) is a global cells, inhibition of proliferation, migration and anchorage- malignancy. Insulin-like growth factor (IGF) signaling axis independent growth following knockdown of SELL plays critical roles for tumorigenesis. This study defined the expression was noted. In SAS HNSCC cells, expression of clinical and functional roles of IGF binding protein-5 exogenous SELL resulted in the increase of invasion, (IGFBP-5) in OSCC. Down-regulation of IGFBP-5 mRNA anchorage-independent growth and the xenographic tumori­ expression was found during the progression from oral genesis in nude mice. Knockdown of FUCA1 and treatment precancerous lesions to OSCC. The down-regulation in of IGFBP6 inhibited the migration of OECM-1 cells. OSCC was associated with a higher propensity to nodal Knockdown of RGS1 inhibited the anchorage-independent metastasis. SAS and OECM-1 are OSCC cells which have growth of SAS cells. Our results provide a useful gene and have not IGFBP-5 expression, respectively. Recombinant signature profile describing the factors underlying the IGFBP-5 reduced the proliferation of OECM-1 cell, and this metastasis of HNSCC to the cervical lymph nodes, which was exerted mainly through blockage of the IGF pathways. may be beneficial for the treatment of HNSCC metastasis. Either IGFBP-5 or IGF-I treatment alone promoted OECM-1 migration, but a combination of treatments generated 51. Initial diagnosis of vitamin B12 deficiency by antagonistic effects. Overexpression of IGFBP-5 reduced sore mouth and assessment of the red blood cell the proliferation and anchorage-independent growth of both dis­tribution width, mean corpuscular volume, and OECM-1 cell and SAS cells. Whereas, knockdown of hemoglobin level - a preliminary study IGFBP-5 expression significantly induced the proliferation Shin-Yu Lu, Ming-Chung Wang and anchorage-independent growth of SAS cells. It also Chang Gung Memorial Hospital Kaohsiung Medical Center, induced the growth of xenografted SAS tumors. SAS Chang Gung University College of Medicine, Kaohsiung, Taiwan transfectants expressed mutant or truncated IGFBP-5, which The oral manifestations of glossodynia, , and lack IGF binding activity exhibited significantly lower stomatitis in vitamin B12 deficiency have long been anchorage-independent growth than vector control. This recognized. These oral changes may occur in the preanemic suggested the existence of IGF-independent suppressor stage. The aim of this paper was to study the red blood function in IGFBP-5. The suppressive effects of IGFBP-5 on cell distribution width (RDW), MCV (mean corpuscular the tumorigenesis of OSCC might be invaluable to future volume), and hemoglobin (Hb) level in 34 patients with neoplastic intervention. a wide range of oral symptoms and signs as the initial manifestations of vitamin B12 deficiency. Of the 34 patients, 53. Association of expression aberrances and 29 were normal to mildly anemic, and 5 were moderately to genetic polymorphisms of lysyl oxidase (LOX) with severely anemic; none had generalized symptoms sufficient areca-associated oral tumorigenesis to arouse suspicion of micronutrient deficiency or anemia Tzong-Ming Shieh1, Shu-Chun Lin1, Chung-Ji Liu1,2, before they visited the Oral Medicine Clinic. Nine (26.4%) Shu-Shin Chang1, Ti-Hsuan Ku1, Kuo-Wei Chang1,3* of the 34 patients had normal RDWs and 8 (88.9%) of these Institute of Oral Biology1, School of Dentistry, National Yang- 9 patients also had normal MCVs. Overall, 13 (38.2%) of Ming University, Taipei, Taiwan; Oral and Maxillofacial the 34 patients had normal MCVs and 3 (8.8%) had low Surgery2, Taipei Mackay Memorial Hospital, Taipei, Taiwan; MCVs. Our findings indicate that the classic presentation of and Department of Dentistry3, Taipei Veterans General Hospital, macrocytic heterogeneous anemia in vitamin B12 deficiency Taipei, Taiwan does not hold true in a large proportion of cases, and the Areca nut use is the major cause of oral squamous cell increase of RDW is not necessarily the earliest indicator of carcinoma (OSCC) in Southern Asians. Areca nut contains a vitamin B12 deficiency. Therefore, a diagnosis of vitamin high level of free copper ions. Lysyl oxidase (LOX) is a B12 deficiency should be considered even in patients with copper-activated enzyme critical for extracellular matrix normal Hb levels and low or normal MCVs. Nevertheless, organization. Contradictory evidence has been put forward Oral Med Pathol 12 (2008) 81 to suggest that LOX may be either an oncogenic or a adjuvant radiotherapy. In conclusion, the MDM2 SNP309 suppressive element. This study investigated the oncogenic and p53 codon 72 SNP may be useful prognostic indicators significance of LOX in areca-associated OSCC. The for patients with OSCC treated with adjuvant radiation. expression assays and polymorphism analysis were performed to know the clinicopathological implications of 55. Areca-induced MMP-2 production in oral LOX status in OSCC. Knock-down and overexpression fibroblast enhances tumorigenesis of oral carcinoma experiments were conducted to know the phenotypic impacts cell of LOX on OSCC cells. Up-regulation of LOX mRNA and Hsuan-Hsuan Lu1, Chung-Ji Liu1,2, Tsung-Yun Liu1,3, LOX protein expression in OSCCs relative to adjacent oral Kuo-Wei Chang1,3* and Shu-Chun Lin1* mucosa were found. Precancerous lesions had the highest Institute of Oral Biology1, School of Dentistry, National Yang- LOX mRNA expression. Areca nut extract up-regulated Ming University, Taipei, Taiwan; Oral and Maxillofacial LOX expression in oral epithelial cells. Knock-down of Surgery2, Taipei Mackay Memorial Hospital, Taipei, Taiwan; LOX induced cellular migration and invasion, but it reduced and Department of Medical Education and Research3, Taipei the anchorage-independent growth and xenographic Veterans General Hospital, Taipei, Taiwan tumorigenesis of OSCC cells. The reduction of migration Areca (betel) nut chewing is tightly linked to oral and invasion by LOX overexpression were partially rescued mucosal and submucosal pathogenesis in Asians. Although by blockage of LOX activity. The Arg158Gln polymorphism there are evidences supporting that the areca nut is a group I was associated with earlier clinical stage of OSCC. Wild carcinogen, the areca nut-associated pathogenesis in oral type LOX overexpression induced anchorage-independence target cells requires further elucidation. This study growth in OSCC cells, but this was not for LOXArg158Gln investigated the effects of areca nut extract (ANE) on overexpression. In conclusion, LOX exerts oncogenic roles primary normal human oral fibroblasts (NHOFs) and the in areca-associated OSCC. This potential could be affected consequential impacts on epithelial carcinogenesis. The by the existence of LOX propeptide domain or genetic genotypic effects of chronic subtoxic ANE treatment in polymorphism. NHOF were examined using RT-PCR and Western blotting. The effects of the supernatant of ANE-treated NHOF and 54. MDM2 SNP 309 G/G and p53 codon 72 Arg/Arg MMP-2 treatment were examined using proliferation, genotypes predict poor survival and control of oral migration and colony formation assays and Western blotting. carcinoma after postoperative irradiation The tumorigenic promoting ability of ANE-treated NHOF Hsi-Feng Tu1,2,5, Shu-Chun Lin1, Hong-Wen Chen3, was examined by subcutaneous co-injection of oral cancer Shou-Yen Kao1,4, Chung-Ji Liu1,2,5* cells with ANE-treated NHOF into nude mice. Chronic Institute of Oral Biology1, School of Dentistry, National Yang- subtoxic ANE treatment resulted in growth inhibition and Ming University, Taipei, Taiwan; Department of Dentistry2, I- the appearance of SA-β-Gal in NHOFs. A significant Lan Hospital, I-Lan, Taiwan; Department of Radiation Oncology MMP-2 activation was elicited in the ANE-induced senescent and Hospice Center3, Taipei Mackay Memorial Hospital, Tai­ (as)NHOFs. Oral cancer cells treated with asNHOF super­ pei, Taiwan; Department of Dentistry4, Veterans General Hos­ natant exhibited enhancement in proliferation, migration and pital, Taipei, Taiwan; and Department of Oral and Maxillofacial anchorage-independent growth. The treatment also dras­ Surgery5, Taipei Mackay Memorial Hospital, Taipei, Taiwan tically activated IGFR and AKT, and up-regulated COX-2 The p53 tumor suppression pathway is important in and cyclin D1 expression. Blockage of MMP-2 and AKT effects associated with radiotherapy. The mouse double significantly attenuated the enhancement in anchorage- minute 2 (MDM2) gene plays a pivotal role in this pathway independent growth. Subcutaneous co-injection of cancer by down-regulating p53. A functional T-to-G polymorphism cells with asNHOFs into nude mice significantly enhanced at nucleotide 309 in intron 1 (SNP309) has been recently tumorigenesis by SAS cells and also induced the transient identified in the MDM2 promoter region. Cells carrying the xenographic tumors from non-tumorigenic OECM-1 cells. SNP309 G allele are reported to have higher MDM2 In conclusions, the senescence stress induced by ANE may transcription activity. We sequenced MDM2 SNP309 and aid oral cancer progression through up-regulating MMP-2 in p53 codon 72 Arg to Pro SNP in 211 patients with oral asNHOFs. These findings support an interactive role for squamous cell carcinoma (OSCC), 70 with oral submucous areca nut-exposed stromal cells in the progression of oral fibrosis, and 116 healthy controls. Neither MDM2 SNP309 cancers among a risk population. nor p53 codon 72 SNP were associated with susceptibility to or the age at onset of OSCC. A higher Arg allelic frequency 56. Aggressive psammomatoid ossifying fibroma in in p53 codon 72 was associated with more advanced disease. the paranasal sinus - case report T, N, and p53 codon 72 SNP were independent predictors of Ming-Hsuan Hsu1, Jyh-Kuei Chen2, Meng-Ling Chiang1, overall (OS) and disease-free survival (DFS). The MDM2 Chi-Ying Tsai2 SNP309 G/G genotype was significantly associated with Department of General Dentistry1, Department of Oral and poor survival in advanced OSCC. The combination of Maxillofacial Surgery2, Chang Gung Memorial Hospital, MDM2 SNP309 G/G and p53 codon 72 Arg/Arg genotypes Chang Gung University College of Medicine, Taipei, Taiwan predicted the worst DFS. This association with poor OS and Aggressive psammomatoid ossifying fibroma (APOF) is DFS was confirmed in the subgroup of patients treated with a rare fibro-osseous lesion found in the head and neck 82 Proceedings of the 3rd Annual Meeting of the Asian Society of Oral and Maxillofacial Pathology region. It arises predominantly within the sinonasal and hyperkeratosis (0.26%), leukoplakia (0.15%), and erythroleu­ orbital in children and young adults. The tumor bears koplakia (0.05%). The most frequent type of recurrent distinctive histomorphological features and has a tendency aphthous ulcer was the minor type (3.42%), followed by the of locally aggressive behavior. The high recurrence rate major type (0.51%), and herpetiform type (0.25%). Burning might be attributed to difficulty in proper resection caused mouth syndrome showed a female-to-male ratio of 1.77: 1 by the location and infiltrative nature of tumor borders. A with xerostomia and dry eye as the most commonly 44-year-old female complained of a large firm mass over the associated symptoms. The prevalence of lichen planus was left side of the hard palate. The periapical films showed a 1.39% and 1.01% in men and women, respectively. It well-demarcated radiolucent lesion extending from the consisted mainly of reticular and erosive types. The current apices of the left upper first premolar to the left upper second study showed a wide spectrum of oral mucosal conditions molar. The root of the left upper second premolar and the left and lesions in a northern Taiwanese population. upper first molar were resorbed. The aspiration cytology revealed a negative result for malignancy while the incisional 58. Central granular cell odontogenic tumor - case biopsy demonstrated numerous tiny round to oval ossicles report resembling psammomatoid bodies with myxomatous matrix. Ting An Chou1, Cheng-Pang Lan1, Meng-Ling Chiang2, It, therefore, was diagnosed as aggressive psammomatoid Yung-Ming Chang3 ossifying fibroma. This patient then was managed with Department of Periodontology1, Department of General Den­ tumor excision and extraction of involved teeth. The tistry2, Department of Oral and Maxillofacial Surgery3, Chang differential diagnosis from other entities is discussed in Gung Memorial Hospital, Chang Gung University College of detail with emphasis on histopathological characteristics. Medicine, Taipei, Taiwan Moreover, the treatment alternatives and prognosis are Central granular cell odontogenic tumor (CGCOT) is a mentioned here as well. rare lesion. Only 30 cases were documented in the literatures. CGCOT is mostly a painless tumor, which is usually located 57. Prevalence of oral mucosal conditions and in the mandibular molar area. A 43-year-old female came to lesions in a northern Taiwanese population the Oral and Maxillofacial Surgery Department with a Yu-Jia Hsieh1, Meng-Ling Chiang1, Chun-Pin Chiang2,3 complaint of a swelling mass over the leftupper area. The Department of General Dentistry1, Chang Gung Memorial swelling mass was incidentally found during the extraction Hospital, Chang Gung University College of Medicine, Taipei, of her left maxillary first molar in a dental clinic in Shanghai. Taiwan; School of Dentistry2 and Dental Department of National On clinical examination, a firm swelling was palpated over Taiwan University Hospital3, College of Medicine, National the left upper molar region. A panoramic radiograph showed Taiwan University, Taipei, Taiwan a radiolucent lesion extending from the left maxillary first The aim of this study was to evaluate the prevalence of premolar to the left maxillary second molar region. The oral mucosal conditions and lesions in a northern Taiwanese tumor was totally excised. Microscopically examination of population. The study group comprised of 1958 consecutive the specimen showed a central granular cell odontogenic patients (866 men and 1092 women) receiving dental tumor. Immunohistochemical study revealed that the tumor examination and treatment in the Department of General cells were positive NSE, vimentin, and CD68, weakly Dentistry, Chang Gung Memorial Hospital starting from positive for S100, and negative for AE1/AE3. 2003 to 2007. The ages of the patients ranged from 4 to 91 (mean, 46±20) years. The most frequently observed oral 59. The EpCAM expression and promoter meth­ mucosal condition was Fordyce’s granules (82.69%), ylation in oral squmaous cell carcinoma followed by the buccal (34.68%), Yi-Shing Shieh1,2, Shine-Gwo Shiah3, Long-Chang (23.70%), (21.14%), commissural lip pit Chang1,2 (0.51%), and cleft lip and/or cleft palate (0.41%). Fordyce’s Department of Dentistry1, Tri-Service General Hospital, Taipei, granules were more common seen in adults than in children. Taiwan; School of Dentistry2, National Defense Medical Center, Buccal exostosis occurred most commonly in the anterior Taipei. Taiwan; and Institute of Cancer Research3, National region. The prevalence of torus palatinus and torus mandibu­ Health Research Institutes, Miaoli, Taiwan laris peaked at the early adult life. The tongue lesions Epithelial cell adhesion molecule (EpCAM) is important observed in the descending order were as follows: lingual for cell proliferation and differentiation. However, its role in varices (16.60%), (1.33%), fissured oral squamous cell carcinoma (OSCC) is not well-established tongue (1.28%), (0.56%), cleft tongue and the mechanisms affecting EpCAM expression are not (0.31%), atrophic glossitis (0.20%), and median rhomboid clear. The present study is designed to examine clinico­ glossitis (0.20%). The prevalence of lingual varices increased pathologic significance of EpCAM expression in OSCCs with age. The strong association has been found between and the possible involvement of DNA methylation machinery and geographic tongue in other studies. in the regulation of EpCAM expression during tumor However, this close association did not show in this study. development and progression. Immunohistochemical stain­ The prevalence of the oral mucosa lesions in the descending ing for the expression of EpCAM and DNA methyltransferase order was recurrent aphthous ulcer (4.34%), burning mouth 1 (DNMT1) was performed in 78 cases of OSCC. Laser syndrome (1.84%), lichen planus (1.71%), frictional microdissection (LMD) was used to obtained cancer cells Oral Med Pathol 12 (2008) 83 from tumor specimens and genomic DNA was extracted in 60. Malignant peripheral nerve sheath tumor of the those cells. Methylation status of the EpCAM promoter was mandible Report of two cases examined by the methylation-specific polymerase chain Ching-Yi Chen, Wen-Chen Wang, Yuk-Kwan Chen, reaction (MSP) assay in 58 OSCC specimens. The resultant Li-Min Lin immunoreactivity and methylation are correlated with Department of Oral Pathology, School of Dentistry, College of clinicopathologic features. EpCAM expression was not Dental Medicine, Kaohsiung Medical University, and Kaohsiung detected in normal epithelium and high expression was Medical University Hospital, Kaohsiung, Taiwan observed in 52.6% (41/78) of OSCC samples. Heterogeneous Malignant peripheral nerve sheath tumors (MPNSTs) and plasticity change of EpCAM expression was noted are almost always found in soft tissue and they have an during tumor development. Of the OSCC samples analyzed, association with neurofibromatosis type 1 (NF-1). The methylation allele was found in 50% (29/58) of cases. We MPNSTs occurring in patients with NF-1 account for 25% to found EpCAM expression was associated with promoter 50% of all observed cases. Primary intraosseous MPNST is methylation significantly (P = 0.032). However, EpCAM a very rare malignant neoplasm. The mandible is the most expression and promoter methylation status didn’t correlate frequently site among the 8 cases that have been reported. with clinicopathologic variables of OSCC. DNMT1 We reported two cases of primary intraosseous MPNST in expression was observed occasionally in basal cells of nor­ the mandible. No metastasis could be identified in both mal epithelium. High expression was observed in 56.4% cases. Segmental resection and flap reconstruction were (44/78) of OSCCs. There was no association between preformed. No local recurrence or metastasis occurred in the DNMT1 and EpCAM expression or EpCAM methylation first case during the follow-up. However, local bone invasion status. Notably, high DNMT1 expression correlated with was suspected in the second case after 5-month follow-up tumor size (P = 0.019) and histologic differentiation (P = and thus chemotherapy was given. MPNSTs may arise from 0.034) of OSCC. Our data implicated that EpCAM expression a pre-existing benign peripheral nerve sheath tumor or de is dynamic change during cancer development of OSCC. novo from a peripheral nerve trunk or following radiation Hypermethylation of EpCAM promoter is associated with therapy. The image, gross and microscopic findings of our EpCAM expression levels in OSCCs, suggesting an both cases showed de novo formation from the right inferior epigenetically mediated loss of EpCAM expression playing alveolar nerve. In case 2, the FDG-PET CT examination a role of its regulation in this tumor. The increased DNMT1 displayed reliable ability for detection of malignant neoplasm protein expression may involve in histogenesis and by comparing the uptake value of FDG avid foci. MPNSTs progression of OSCC. are difficult to diagnose due to their rarity and variation. However, the diagnosis is usually dependent on microscopic findings and required immunohistochemical staining.