associated with cancer cell invasion and tumor immunity. The aim of this study is to investigate the clinical significance of expression of AhR in RCC and infiltrating lymphocytes. Furthermore, we elucidate the role of AhR in cancer cells by regulating the expression using its ligand or siRNA for AhR in vitro. METHODS: A total of 120 RCC patients were included in the study. The median age was 60 years (25 to 87) and the median follow-up was 63 months (0.3 to 151). The AhR expression in cancer cells and infiltrating lymphocytes in stromal tissues adjacent to cancer nests was investigated by immunohistochemistry and its associations with the clinicopathological parameters and prognosis were analyzed. AhR signal pathway was activated by exposure to 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD), a ligand of AhR, or AhR expression was down-regu- lated by siRNA in an RCC cell line, 786-O. Subsequently, cancer cell invasion and expression of MMPs were investigated using Matrigel TM and real-time quantitative PCR. RESULTS: The AhR expression in RCC and infiltrating lym- phocytes was significantly higher in clear cell RCC compared to non- clear cell RCC (p < 0.05). Therefore, further experiments were confined to clear cell RCC. AhR in RCC was associated with the smoking history (p ¼ 0.007), gender (p ¼ 0.002) and histological grade (p ¼ 0.02). The number of AhR-positive lymphocytes was significantly associated with AhR expression in RCC (p < 0.01), pathological T (p < 0.05), patho- logical N (p < 0.05), vascular invasion (p < 0.01) and metastasis (p < 0.05). A multivariate analysis revealed that the histological grade (p < 0.001), vascular invasion (p < 0.01) and AhR in cancer cells (p < 0.001) were independent prognostic factors for the PFS. The 5-year PFS rates were 80.5% in the low AhR RCC and 20.4% in the high AhR RCC (p < 0.001). TCDD treatment enhanced the cell invasion through Matrigel TM (p < 0.05) together with up-regulating the expression of MMP-1 and MMP-9 (p < 0.05). The cell invasion through Matrigel TM was inhibited in comparison to the cells transfected with a non-targeting siRNA (p < 0.05) together with the down-regulation of MMP-1 and MMP-9 (p < 0.05). CONCLUSIONS: These results suggest that AhR in cancer cells regulates invasion of RCC and that AhR may be involved in tumor immunity. AhR could therefore be a potentially attractive therapeutic target for the control of RCC. Source of Funding: Grant-in-Aid for Young Scientists (Start up) (No. 21890250) (M.I.), Grant-in-Aid for Young Scientists (B) (No. 24791668), Grant-in-Aid for Scientific Research (C) (No.25460422) (S.M.), Grants-in-Aid for Scientific Research (S) (No. 19109004) (Y.O.), Grain-in-Aid for Scientific Research (B) (No. 24390374) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) (M.O.) and Project for Development of Innovative Research on Cancer Therapeutics (P-Direct) (S. M., R.M, K.T. and M.O.) from MEXT MP29-03 TET3, HELLS, TOP2A AND ATAD2 ARE NOVEL INDEPENDENT PROGNOSTIC MARKERS IN ADVANCED RENAL CELL CARCINOMA Dong Chen*, Munich, Germany; Matthias Maruschke, Rostock, Germany; Rainer Riesenberg, Wolfgang Zimmermann, Christian. G. Stief, Alexander Buchner, Munich, Germany INTRODUCTION AND OBJECTIVES: A significant proportion of patients with renal cell carcinoma (RCC) will have metastases at first diagnosis or develop metastases during the course of their disease. Individual risk stratification based on reliable prognostic markers is needed to allow a personalized therapeutic strategy. The aim of our study was to screen for promising prognostic markers using a novel filtering approach, based on genes that are subsequently up-regulated in primary tumors and metastases. Therefore, we created microarray expression data from RCC patients including normal kidney tissue, primary tumors and metastases. The best candidate genes were vali- dated using RT-PCR. METHODS: Tissue samples from normal renal tissue (n¼14), primary tumor (14x G1, 14x G3) and metastases (n¼32) were collected from patients with clear-cell RCC. Follow-up data were available from all patients. The tissue was snap-frozen immediately after surgery, and total RNA was isolated. Expression profiles were created using oligo- nucleotide microarrays (GeneChip HG U133 Plus 2.0, Affymetrix). For analysis of expression data, the dChip 2010 software was used. Gene expression was compared between normal kidney, primary tumor and metastases. Furthermore, a significance test was applied. In all filtered genes, univariate survival analysis was carried out (Kaplan-Meier method, log-rank test). Significant genes were further analyzed using multivariate Cox regression models. The best candidate markers were further validated in an independent cohort of 52 primary tumors using quantitative RT-PCR and immunohistochemistry. RESULTS: Expression analysis revealed that in 59 probe sets there was increased expression in primary RCC compared with normal kidney, and also in metastases compared with primary tumors. In uni- or multivariate survival analysis, 15 of these probe sets were significant. Undefined genes (C1orf216 and Hs.133294.1) and genes with very low mRNA level in RT-PCR (NDC80, GOLSYN, DTL, and BUB1B) were excluded. Eight genes (TOP2A, SFN, CENPF, AMPD3, ATAD2, AURKA, HELLS and TET3) were afterwards validated by quantitative RT-PCR. Survival analysis in an independent cohort of 52 primary RCCs showed that TOP2A (HR¼4.4, p¼0.004), TET3 (HR¼2.9, p¼0.031), HELLS (HR¼3.6, p¼0.007), and ATAD2 (HR¼3.8, p¼0.014) represent independent predictors for RCC patients. TOP2A IHC results support the RT-PCR findings. CONCLUSIONS: High expression of TET3, HELLS, TOP2A, and ATAD2 are independent predictors of poor outcome in RCC pa- tients and may be used for individual risk-adapted therapy in the future. Source of Funding: none MP29-04 GAIN OF CHROMOSOME 8Q IS ASSOCIATED WITH INFERIOR SURVIVAL IN PATIENTS WITH RENAL CELL CARCINOMA Reza Mehrazin*, Robert G. Uzzo, Essel Dulaimi, Karthik Devarjan, Jeffrey J. Tomaszewski, Joseph Testa, Jianming Pei, Philip Abbosh, Timothy Ito, Alexander Kutikov, Tahseen Al-Saleem, Philadelphia, PA INTRODUCTION AND OBJECTIVES: The proto-oncogene c-MYC, located on chromosome 8q, can be up-regulated through gain of 8q causing alteration in biology of renal cell carcinoma (RCC). The aim of this study was to evaluate the prevalence of chromosome 8q gain in our renal cancer tumor registry and to correlate findings with tumor histology, grade, cancer specific survival (CSS), and overall survival (OS) in clear and papillary METHODS: Cytogenetic analysis by conventional or Chromo- somal Microarray Analysis (CMA) was performed on tumors of 414 patients who underwent radical or partial nephrectomy for suspected RCC. Non-clear RCC, non-papillary RCC, and those which did not show alteration or gain in chromosome 8 on cytogenetic analysis were excluded from the study. Impact of gain in chromosome 8q status on CSS, OS, and its correlation with clinicopathological variables, obtained from our prospectively maintained kidney cancer database, were eval- uated and compared to RCC’s that did not have gain in chromosome 8q. CSS and OS were assessed using log-rank test and the Cox pro- portional hazards model. RESULTS: After the exclusions total 279 clear cell and papillary RCC tumors with cytogenetic analysis were included in study. Gain of 8q was detected in 18 (6.5%) tumors (9 clear cell, 3 sarcomatoid variant, and 6 papillary RCC), using either conventional method (11) or CMA (7). Gain of 8q was associated with higher grade (12% vs 2.0%, p ¼ 0.001), higher risk of regional lymph node involvement (30 % vs. 6%, p¼0.008), and distant metastasis (22 % vs. 4.0%, p<0.001). Among tumors with 8q gain, 56% were stage III or higher. No Vol. 191, No. 4S, Supplement, Sunday, May 18, 2014 THE JOURNAL OF UROLOGY â e305