RGC-32参与TGF-β#35825;导肾小管上皮向间充质细胞转化的分子调控机制

项目名称： RGC-32参与TGF-β#35825;导肾小管上皮向间充质细胞转化的分子调控机制

项目编号： No.30871177

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 黄文彦

作者单位： 复旦大学

项目金额： 32万元

中文摘要： 肾小管上皮-间充质细胞转化(EMT)在肾间质纤维化发生发展中发挥重要作用，TGF-β#20026;EMT最重要的诱导因子，但其确切的分子调控机制尚远不清楚。晚近我们的工作提示Response Gene to Complement 32(RGC-32)为TGF-β#20449;号激活通路下游关键的转录调控因子，但其参与调控EMT的分子机制不详。本研究在体外培养正常大鼠肾小管上皮细胞(NRK-52E)基础上，(1)利用基因克隆，基因转染，凝胶电泳迁移分析(EMSA)，染色体免疫共沉淀等技术，明确TGF-β#35825;导EMT过程中，与TGF-β#30456;关的RGC-32基因启动子及其调控元件。(2)利用阻断已知TGF-β#19979;游信号通路，过表达和抑制RGC-32等手段，初步了解RGC-32参与调控EMT可能的信号通路以及信号通路间的串话调节。从而为阐明TGF-β#20171;导EMT的分子机制提供理论依据。

中文关键词： 肾间质纤维化; EMT; RGC-32; 基因调控; 信号转导

英文摘要： It is widely accepted that epithelial to mesenchymal transition (EMT) is a central mechanism for the tubular interstitial fibrosis (TIF), while the transforming growth factor-βTGF-β and its downstream Smad signaling play an essential role in EMT. We recently identified that Response Gene to Complement 32 (RGC-32) to be critical gene for TGF-βinduced EMT; however, the molecular regulation mechanisms were so far documented. Based on culture of rat renal tubular epithelial cell lines (NRK-52E) in vitro, this study will be (1) to identify the related promoters and their regulation elements of RGC-32 gene in TGF-βinduced EMT, using gene cloning, gene transfection, Electrophoretic Mobility Shift Assays (EMSAs), and chromosome CHIP technology. (2) to understand the possible signaling transductions and their key regulation cross-talking factors in TGF-βinduced EMT, using blocking the downstream signaling of TGF-βsuch as Smads signaling transduction, Rho A, et al ) and gene cloning technology(gene transfection, siRNA, and antibody) to over-expression or down-expression of RGC-32.

英文关键词： Tubular interstitial fibrosis; EMT;RGC-32; Gene Regulation; Signaling Transduction