抑癌基因ECRG4在肾癌中的表达调控及功能研究

项目名称： 抑癌基因ECRG4在肾癌中的表达调控及功能研究

项目编号： No.81201579

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 肿瘤学1

项目作者： 钱学敏

作者单位： 深圳大学

项目金额： 23万元

中文摘要： 抑癌基因ECRG4异常甲基化与多种肿瘤的发生发展密切相关。但是，该基因与肾癌的关系尚无报道；也未见有关组蛋白修饰情况对ECRG4基因表达影响的研究。本项目前期工作及预实验结果显示：ECRG4基因在正常肾组织中高表达，但在肾癌细胞株和肾透明细胞癌组织中表达显著下调，且表达量与其启动子甲基化程度成负相关。免疫组化染色也显示ECRG4蛋白在癌组织中表达比癌旁正常组织低。因此，我们推测ECRG4在肾癌中是个抑癌基因。为此，本课题组拟：(1)采用RT-PCR、Western blot和免疫组化等技术，分析ECRG4基因表达及其与肾癌临床病理特征的相关性；（2 使用亚硫酸氢盐基因组测序和染色质免疫共沉淀等技术分析ECRG4基因表达的表观调控机制；（3）利用慢病毒表达系统等技术研究ECRG4基因的功能。以期初步揭示ECRG4在肾癌发生中的作用，为肾癌的早期辅助诊断和基因治疗提供新依据。

中文关键词： 肾癌；预后；ECRG4；免疫组化；DNA甲基化

英文摘要： The promoter hypermethylation of novel candidate tumor suppressor gene ECRG4 is closely related with the development of a varity of tumors. However, expression and role of ECRG4 in renal cell carcinoma have not yet been studied, and effects of histone modification on ECRG4 expression have not yet been reported. Our previous work and preliminary results show that ECRG4 is highly expressed in normal renal tissue but is significantly reduced in renal carcinoma cell lines and renal clear cell carcinoma, and that its expression is negatively correlated with its promoter methylation levels. Immunohistochemical staining has demonstrated that ECRG4 protein in cancerous tissues is lower than that in adjacent normal tissues.Therefore, we come up with a hypothesis that ECRG4 is a candidate tumor suppressor gene in renal cell carcinoma. To attain this goal , we will investigate (1) ECRG4 expression and its association with linicaopathologicalparameters in clinical samples by using real-time quantitative PCR, Western blot and immunohistochemistry;(2) its epigenetic regulation mechanisms of ECRG4 in renal cell carcinoma by bisulfite genomic sequencing and chromatin immunoprecipitation; (3) the function and role of ECRG4 in renal cell carcinoma by lentiviral vector system. All these investigations are to provide new clues f

英文关键词： renal cancer；prognosis；ECRG4；immunohistochemistry；DNA methylation