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Gene Products of Chromosome 11Q and Their Association with CCND1

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Gene Products of Chromosome 11Q and Their Association with CCND1 Available online http://breast-cancer-research.com/content/10/5/R81 ResearchVol 10 No 5 article Open Access Gene products of chromosome 11q and their association with CCND1 gene amplification and tamoxifen resistance in premenopausal breast cancer Katja Lundgren1, Karolina Holm2, Bo Nordenskjöld3,4, Åke Borg2 and Göran Landberg1 1Center for Molecular Pathology, Department of Laboratory Medicine, Lund University, Malmö University Hospital, Malmö, SE-205 02, Sweden 2Department of Clinical Sciences, Division of Oncology, Lund University, Lund, SE-221 85, Sweden 3Department of Oncology, Borås Hospital, Borås, SE-501 82, Sweden 4Division of Oncology, Faculty of Health Sciences, Linköping University, Linköping, SE-581 85, Sweden Corresponding author: Göran Landberg, [email protected] Received: 2 Apr 2008 Revisions requested: 8 May 2008 Revisions received: 4 Aug 2008 Accepted: 29 Sep 2008 Published: 29 Sep 2008 Breast Cancer Research 2008, 10:R81 (doi:10.1186/bcr2150) This article is online at: http://breast-cancer-research.com/content/10/5/R81 © 2008 Lundgren et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Introduction The amplification event occurring at chromosome protein expression was also compared with gene expression locus 11q13, reported in several different cancers, includes a data in a subset of 56 breast cancer samples. number of potential oncogenes. We have previously reported amplification of one such oncogene, namely CCND1, to be correlated with an adverse effect of tamoxifen in premenopausal Results Cortactin and FADD (Fas-associated death domain) breast cancer patients. Over-expression of cyclin D1 protein, over-expression was linked to CCND1 amplification, however, confers tamoxifen resistance but not a tamoxifen- determined by fluorescence in situ hybridization, but was not induced adverse effect. Potentially, co-amplification of an associated with a diminished effect of tamoxifen. However, additional 11q13 gene, with a resulting protein over-expression, deletion of distal chromosome 11q, defined as downregulation is required to cause an agonistic effect. Moreover, during 11q13 of the marker Chk1 (checkpoint kinase 1), was associated with amplification a deletion of the distal 11q region has been an impaired tamoxifen response, and interestingly with low described. In order to assess the potential impact of the deletion proliferative breast cancer of low grade. For Pak1 (p21- we examined a selected marker for this event. activated kinase 1) and cyclin D1 the protein expression corresponded to the gene expression data. Method Array comparative genomic hybridization analysis was employed to identify and confirm changes in the gene expression of a number of different genes mapping to the 11q Conclusions The results indicate that many 11q13 associated chromosomal region, associated with CCND1 amplification. gene products are over-expressed in conjunction with cyclin D1 The subsequent protein expression of these candidate genes but not linked to an agonistic effect of tamoxifen. Finally, the was then examined in a clinical material of 500 primary breast deletion of distal 11q, linked to 11q13 amplification, might be an cancers from premenopausal patients who were randomly important event affecting breast cancer outcome and tamoxifen assigned to either tamoxifen or no adjuvant treatment. The response. Introduction Amplification of chromosome locus 11q13 occurs at high fre- Gene amplification is a well defined cause of oncogene acti- quencies in certain human cancers, including lung, bladder, vation during tumor development, and some genomic regions breast and ovarian carcinomas, as well as in head and neck are recognized to be more frequently amplified than others [1]. squamous cell carcinomas (HNSCCs) [2-6]. Approximately 15% of primary breast cancers are affected by this specific amplification, which is associated with poor prognosis [7-10]. CISH: chromogenic in situ hybridization; ER: estrogen receptor; CGH: comparative genomic hybridization; Chk1: checkpoint kinase 1; FADD: Fas- associated death domain; FISH: fluorescence in situ hybridization; HNSCC: head and neck squamous cell carcinoma; ICC: immunocytochemistry; IHC: immunohistochemical; LOH: loss of heterozygosity; PR: progesterone receptor; RFS: recurrence-free survival. Page 1 of 14 (page number not for citation purposes) Breast Cancer Research Vol 10 No 5 Lundgren et al. Four distinct core regions or amplicons within the 11q13 deleted in the 11q13 amplification event might also affect locus have been identified, and these can be amplified inde- breast cancer outcome and treatment response. pendently or concurrently in various combinations [7,11]. A number of oncogenes or potential cancer-related genes have In order to elucidate the importance of 11q-associated genes been mapped to the 11q13 chromosomal region. The with regard to tamoxifen response and CCND1 amplification, CCND1 and CTTN oncogenes have been putatively pro- we identified previously described candidate genes at the 11q posed as candidate genes for the emergence and mainte- chromosomal region for further investigation, using array com- nance of this amplification event in breast cancer [3,7]. These parative genomic hybridization (CGH) analysis of breast can- genes map to two different amplification cores, located within cer samples. Protein expression levels of the various cancer- 0.8 megabases of each other at 11q13.3 [2,11], and their co- related gene products were then analyzed in a tissue microar- amplification has been reported in breast cancer [1,2,12]. The ray from a randomized trial of premenopausal breast cancer core region comprising CCND1 is the most frequently ampli- patients receiving 2 years of adjuvant tamoxifen treatment or fied and is involved in two-thirds of the amplifications. The no adjuvant treatment. By comparing treated and untreated CCND1 gene is the most extensively studied gene of the patients, we were able to delineate the response to tamoxifen 11q13 amplification region, and encodes the cell cycle regu- irrespective of prognostic features, and a potential agonistic or latory protein cyclin D1, which is important both for develop- diminished effect of tamoxifen could be identified. Further- ment of mammary tissue and in mammary carcinogenesis [2]. more, the tumor material allowed us to study prognostic fea- tures, relations with different clinicopathological parameters, In breast cancer, amplification and over-expression of cyclin and associations with CCND1 amplification, in different sub- D1 has been associated with worse prognosis [13,14], but groups defined by the expression of the 11q13 and distal 11q high expression of cyclin D1, in contrast, has also been asso- gene products. The results indicate that many 11q13-associ- ciated with better prognosis [15,16]. Breast cancer patients ated gene products are over-expressed in conjunction with exhibiting estrogen receptor (ER)-α expression, with concur- cyclin D1 but are not linked to an agonistic effect of tamoxifen. rent over-expression of cyclin D1, have been reported not to Conversely, deletion of the distal end of chromosome 11q, benefit from treatment with the selective estrogen receptor defined as downregulation of the marker Chk1 (checkpoint modulator tamoxifen, which is in contrast to the evident kinase 1), was associated with an impaired tamoxifen response in ER-α-positive breast cancers with moderate and response, and with low proliferative breast cancer of low low cyclin D1 expression [15]. Jirstrom and coworkers grade. reported that CCND1 amplification was associated with a potential agonistic effect of tamoxifen in ER-α-positive pre- Materials and methods menopausal breast cancer patients, even when not accompa- Comparative genomic hybridization nied by protein over-expression [17]. Array CGH was performed essentially as was previously described [24]. Raw data and normalized data are available INT2, FADD, PAK1, and EMSY are other candidate genes through National Center for Biotechnology Information Gene reported to be included in the 11q13 amplicon [7] and thier Expression Omnibus [GEO: GSE12759]. amplification or protein over-expression has been associated with a poor prognosis in various cancers [1,2,8,9,18-20]. Patient materials Between 1986 and 1991, a total of 564 premenopausal It has been reported that in HNSCC amplification of 11q13 breast cancer patients with invasive stage II disease were involves a loss of distal chromosome 11q (from 11q14.2 to enrolled in a Swedish trial (SBII:2a), in which they were ran- 11qter) through a breakage-fusion-bridge cycle mechanism domly assigned to 2 years of adjuvant tamoxifen (n = 276) or [21]. In this process, genes with important roles in, for no adjuvant treatment (control; n = 288). The aim of the origi- instance, the DNA damage response are lost in the deletion nal study was to compare 2 years of tamoxifen treatment (20 step preceding the amplification. Frequent allelic deletions at or 40 mg/day) versus no adjuvant treatment. Patients were chromosome 11q24–q25 have been reported in both breast included irrespective of hormone receptor status. All patients and ovarian cancer and have been associated with a worse were followed up for recurrence-free survival (RFS) and overall clinical
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