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Gynecologic Tumor Invasion ANNUAL MEETING ABSTRACTS 175A Conclusions: The upregulated expression of these 4 transcription factors in carcinoma 811 Lack of Association between Epidermal Growth Factor Receptor from radical prostatectomy specimens suggests that androgen receptor activation Overexpression and Disease Recurrence in Clear Cell Renal Cell occurs in hormone naïve primary prostate cancer. Interestingly, the increased mRNA Carcinoma expression for SHP in HGPIN suggests it may play a role in prostate cancer initiation M Zhou, J Finke, R Bukowski, R Tubbs. The Cleveland Clinic Foundation, Cleveland, and early progression. Further studies of the expression of these genes at different OH. stages of prostate cancer are planned to determine their relative in initiation and Background: The prognostic significance of epidermal growth factor receptor (EGFR) progression of prostate cancer. overexpression in clear cell renal cell carcinoma (CCRCC) is controversial. Some studies have shown it is an unfavorable prognostic factor, while other studies found 809 Potential Roles of Cytologic T-Lymphocytes and Nature Killer Cells no association between EGFR overexpression and prognosis. Recent reports that anti- in Prostate Basal Cell Layer Disruptions and Tumor Invasion EGFR therapy was ineffective for CCRCC further raised the question about the biological C Zhao, C Mannion, YG Man. Armed Forces Institute of Pathology, Washington, function of EGFR in CCRCC. We studied EGFR overexpression in a cohort of CCRCC DC; Hackensack Medical Center, Hackensack, NJ. with long-term follow-up (> 10 years). Background: The physical disruption of the basement membrane and basal cell Design: 44 CCRCC were used to construct a tissue microarray (TMA). All patients layer is a pre-requisite for prostate tumor invasion. The disruption of the basement were treated with radical/partial nephrectomy and had been followed for >=10 years. membrane is believed to result from elevated proteolytic enzymes, while the mechanism Eleven had cancer recurrence within 1 year, 22 within 1-10 years, and 11 were of basal cell layer disruptions remains elusive. As our previous studies with antibodies recurrence-free more than 10 years aftersurgery. The TMA was stained with anti- to basal cell specific antigens and leukocyte common antigen (LCA) revealed, focal EGFR antibody. EGFR overexpression was scored using two methods. 1. Simple basal cell layer disruptions are consistently surrounded by or adjacent to LCA positive scoring- strong circumferential membranous stain (3), weak circumferential cells (Man et al, Cancer Detect Prev, In press). This current study attempts to assess membranous stain (2), weak partial membranous stain (1) and negative staining (0). whether these cells belong to a cytotoxic cell population. Only staining intensity 2 and 3 were considered positive for EGFR overexpression. Design: Consecutive tissue sections from human prostate tumors (n=30) with co- 2. Composite scoring-the staining intensity (0-3) was multiplied by the percentage of existing normal, hyperplastic, in situ, and invasive components were double positive cells with EGFR overexpression score ranging from 0 to 300. immunostained with two different chronogens for the high molecular weight Results: Using the simple scoring method, EGFR overexpression was found in 7/11 cytokeratin 34bE12 to identify basal layer disruptions, and each of the following (63.6%), 18/22 (81.8%) and 5/11 (45.5%) of patients with recurrence within 1, 1-10 markers, CD4, CD8, CD56, microphage, perforin, and mast cell tryptase, to elucidate the and >10 years after surgery (p>0.05 ). There was no correlation between EGFR correlation of these molecules with focal basal layer disruptions. overexpression and tumor recurrence status or tumor stage (p=0.37 and 0.55). Results: Multiple focal basal cell layer disruptions were seen in each of the cases. However, EGFR overexpression correlated with Furhman nuclear grade (r=0.372, These disruptions were consistently surrounded by or adjacent to CD8, CD56, p=0.036). Using the composite scoring method, mean EGFR overexpression score was perforin, and mast cell tryptase positive cells. Tumor cells overlying focally disrupted 120+78, 122+77 and 82+85 for patients with recurrence within 1, 1-10 and >10 years basal cell layers often displayed distinct morphological alterations in cellular density after surgery respectively (p>0.2). There was no correlation between EGFR and polarity, as well as the nuclear size and shape, compared to the adjacent cells overexpression level and disease recurrence status, tumor stage or Furhman nuclear within the same duct, but away from the disruption. The CD4 and microphage grade. positive cells also appeared to be associated with basal cell layer disruptions, but the Conclusions: EGFR overexpression correlates with Furhman nuclear grades in number of these cells varied substantially and the association was less consistent. CCRCC. However, it is not significantly different between patients with recurrence Conclusions: The consistent detection of CD8, CD56, perforin, and mast cell tryptase within 1, 1-10 and >10 years after surgery. There is no correlation between EGFR positive cells near focal basal cell layer disruptions suggest that cytotoxic T overexpression and prognosis. lymphocytes, nature killer cells and mast tryptase positive cells are likely to promote basal cell layer disruptions and tumor invasion. The development of specific agents to target these cells may have significant clinical value in treatment and prevention of Gynecologic tumor invasion. This study was supported in part by grants DAMD17-01-1-0129 and DAMD17- 812 Prognostic Significance of Peritumor Lymphatic Vessel Density 01-1-0130 from Congressionally Directed Medical Research Programs to Yan-Gao in Early Stage Squamous Cell Carcinoma of the Cervix Man, MD., PhD. G Acs, X Xu, T Pasha, C Chu, PJ Zhang. University of Pennsylvania Medical Center, Philadelphia, PA. 810 Interferon Signal Transduction Pathway Is Disturbed in a Subset Background: The role of angiogenesis in the development and progression of cervical of High Grade Invasive Urothelial Carcinomas carcinoma is established. However, in cervical cancers the earliest feature of M Zhou, P Sadhukhan, R Rackley, J DiDonato. The Cleveland Clinic Foundation, disseminated disease is regional lymph node involvement. Despite its role in tumor Cleveland, OH. dissemination, little is known about the role of tumor lymphangiogenesis in metastases Background: Interferons are known to have a variety of biological activities including and whether lymphatic spread occurs via pre-existing lymphatic channels or vessels antitumor, antiviral and immunomodulatory effects. Recent cell culture and gene newly formed by lymphangiogenesis. The recently developed monoclonal antibody expression profiling studies have suggested that interferon signaling pathways may D2-40 was reported to be a selective marker for lymphatic endothelium useful in play a key role in the development and progression of high grade urothelial carcinoma. identifying lymphatic invasion in various malignant neoplasms. This study evaluated the expression of several key genes in the interferon signaling Design: We examined the intra- and peritumor lymphatic vessel density (LVD) in a pathways in low grade non-invasive (LGTCC) and high grade invasive urothelial series of 112 FIGO stage I and II cervical squamous cell carcinomas using D2-40 carcinoma (HGTCC). immunohistochemistry on formalin-fixed paraffin-embedded tissue sections. The Design: Construction of tissue microarray: 33 invasive HGTCC, 6 LGTCC and 3 lymphatic vessel density within the tumors (intratumor LVD) and within 2 mm of benign urothelium. Immunohistochemical staining and evaluation: Immunostains were the edge of the tumors (peritumor LVD) was determined in 10 high power fields performed with antibodies against NFkB (P50), Stat3, Phospho-Stat3, and ISGF-3g (X400) with the highest number of D2-40 positive lymphatic vessels. The intra- and (p48). Benign urothelium served as control. Stains significantly higher or lower than peritumor LVD was correlated with clinicopathologic tumor features, D2-40 that of benign urothelium were considered as positive or negative for that marker. A immunoreactivity in tumor cells and patient outcome. case was scored as negative only if all 3 tissue cores from the same case were negative. Results: Intra- and peritumor LVD was significantly higher compared to benign A case was scored as positive if any of 3 tissue cores was positive. For NFkB and Stat3, squamous cervical mucosa (p<0.0001). Peritumor LVD (9.56 ± 0.47, mean ± SEM) the cytoplasmic staining was evaluated. For phospho-Stat3, the nuclear staining was was significantly higher compared to intratumor LVD (7.84 ± 0.49) (p<0.01). High evaluated. For P48, both cytoplasmic and nuclear staining was evaluated. peritumor, but not intratumor, LVD was significantly associated with low D2-40 Results: Normal urothelium was negative for NF kB (P50) and Stat3 expression, but immunoreactivity of tumor cells (p=0.005), presence of lymphatic invasion showed consistent positive nuclear stain for phospho-Stat3 and both cytoplasmic (p<0.0001), nodal metastasis (p=0.026) and FIGO stage (p=0.032). Intra- and and nuclear staining for P48. The expression of P50, Stat-3, Phospho-Stat3 and P48 peritumor LVD showed no correlation with patient age, tumor size or grade. High was summarized in the Table. Expression of phospho-Stat3 and P48 was heterogeneous peritumor, but not intratumor,
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