染色质免疫沉淀技术分析HePG2细胞TCF7L2蛋白与IDE基因转录启动子的结合

摘要

TCF7L2 is the key transcriptional factor regulating glucose metabolism through the Wnt signaling pathway. Insulin-degrading enzyme (IDE) plays a key role in insulin degradation in celluar level and is closely related to hyperglycemia, insulin resistance as well as hyperinsulinemia in patients with Type 2 diabetes mellitus (T2DM). To identify the region in IDE gene promoter where the transcriptional factor TCF7L2 can bind in HePG2 cells. Chromatin immunoprecipitation and PCR were performed using the antibody specific for TCF7L2 to verify the binding of TCF7L2 to IDE promoter. Results indicated that IDE gene specific fragments were detected in the DNA fragments immunoprecipitated by antibody specific for TCF7L2. Thus it proved that TCF7L2 protein could bind to the promoter regions of IDE gene in HePG2 cells and may involved in regulation of IDE gene expression.%TCF7L2是一种重要的转录因子,通过Wnt信号途径,调节葡萄糖代谢.胰岛素降解酶(IDE)是细胞水平催化胰岛素降解的最关键的酶,与2型糖尿病(T2DM)高血糖、胰岛素抵抗、高胰岛素血症密切相关.为了检测HePG2细胞内转录因子TCF7L2与IDE基因启动子区的结合情况,采用染色质免疫沉淀技术结合PCR技术检测IDE基因启动子序列.结果表明,在特异性TCF7L2抗体免疫沉淀的DNA片段中扩增出IDE基因启动子序列,因此证实在HePG2细胞内,TCF7L2蛋白可与IDE基因转录启动子的特异区域结合,进而可能参与IDE基因的表达调控.