Identification of Genes Involved in Bladder Cancer

描述（由申请人提供）：项目概述：膀胱移行细胞癌（TCC）在发达国家的所有癌症中发病率排名第4，但其起源和进展的机制仍知之甚少。也有一些有用的诊断或预后的生物标志物为这种疾病。这项建议的长期目标是确定TCC发展早期和晚期步骤中涉及的分子机制。此外，我们希望发现新的生物标志物，以检测，诊断和预后的TCC。为了实现这一点，我们将使用膀胱癌的小鼠模型，其中SV 40大T抗原（SV 40 T）在尿斑蛋白启动子的控制下表达，该启动子将表达限制于膀胱尿路上皮。这些小鼠（UPII-SV 40 T）发展TCC，其之前是类似于人CIS的状况。这一建议的假设是，有差异的基因表达谱和基因调控网络之间的侵袭性TCC和CIS和CIS和遥远的正常出现尿路上皮，这些变化是恶性状态的指示。这一假说是基于多步骤致癌的概念。我们的假设的一个延伸是，CIS是正常尿路上皮和侵袭性TCC之间的中间步骤，应该有相应于这个中间状态的基因表达的变化。具体目标是：1.使用Affymetrix GeneChip（r）Arrays识别非转基因小鼠和UPII-SV 40 T小鼠（CIS和TCC）的正常尿路上皮之间差异表达的基因和基因调控网络。将通过评估患者TCC和正常尿路上皮标本中人类同源物的表达来验证单个基因与人类TCC的同源性。将进行微阵列数据的计算机模拟分析，包括生成基因表达谱（聚类分析）和基于差异表达基因的启动子序列开发启动子模型。2.通过在人膀胱来源细胞系中稳定过表达或siRNA介导的抑制验证基因，评估这些基因在体内的功能和与膀胱癌发生的潜在相关性。相关性：这些研究的完成将促进我们对TCC发生的分子机制的理解。此外，鉴定正常尿路上皮、CIS和TCC之间差异表达的基因将为这种疾病提供急需的诊断和预后标志物。

DESCRIPTION (provided by applicant): Project Summary: Transitional cell carcinoma of the bladder (TCC) ranks 4th in incidence of all cancers in the developed world, yet the mechanisms of its origin and progression remain poorly understood. There are also few useful diagnostic or prognostic biomarkers for this disease. The long-term aim of this proposal is to determine molecular mechanisms involved in early and late steps in the development of TCC. In addition we hope to identify new biomarkers for detection, diagnosis and prognosis of TCC. To accomplish this we will use a mouse model for bladder cancer in which the SV40 large T antigen (SV40 T) is expressed under the control of the uroplakin promoter, which restricts expression to the bladder urothelium. These mice (UPII- SV40T) develop TCC, which is preceded by a condition resembling human CIS. The hypothesis underlying this proposal is that there are differences in gene expression profiles and gene regulatory networks between invasive TCC and CIS and between CIS and distant normal appearing urothelium, and that these changes are indicative of the malignant state. This hypothesis is based on the multistep carcinogenesis concept. An extension of our hypothesis is that CIS is an intermediate step between normal urothelium and invasive TCC and that there should be changes in gene expression corresponding to this intermediate state. The specific aims are: 1. Identify the genes and gene regulatory networks that are differentially expressed between the normal urothelium of nontransgenic and UPII-SV40T mice, CIS and TCC, using Affymetrix GeneChip(r) Arrays. The relevence of individual genes to human TCC will be validated by assessing expression of the human homologs in patient TCC and normal urothelium specimens. In silico analysis of the microarray data, which includes generating gene expression profiles (clustering analysis) and the development of promoter models based on the promoter sequences of the differentially expressed genes will be performed. 2. Assess the function and potential relevance to bladder carcinogenesis of these genes in vivo by stable overexpression, or siRNA-mediated suppression, of the validated genes in human bladder-derived cell lines. Relevence: Completion of these studies should advance our understanding of the molecular events underlying TCC development. In addition, the identification of genes differentially expressed between normal urothelium, CIS and TCC will provide much needed diagnostic and prognostic markers for this disease.