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.pdfVIP

16 卷 1 期 生命科学研究 灾燥造援员6 晕燥援1 圆园12 年2 月 蕴蚤枣藻 杂糟蚤藻灶糟藻 砸藻泽藻葬则糟澡 Feb. 圆园12 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 * 许丹, 任 伟, 郑晓雅, 毕健琨 （重庆医科大学附属第一医院 内分泌科, 中国重庆400016 ） 摘 要: 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冤是细胞水 平催化胰岛素降解的最关键的酶, 与2 型糖尿病 T2DM冤高血糖尧 胰岛素抵抗尧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. 关键词: 染色质免疫沉淀曰TCF7L2 蛋白曰IDE 基因 中图分类号: R587.1 文献标识码: A 文章编号: 1007 7847(2012)01 0054 05 Detection of TCF7L2 Binding to IDE Gene Promoters by Chromatin Immunoprecipitation * XU Dan, REN Wei , ZHENG Xiao ya, BI Jian kun （Dep artment of Endocrinology, First Affil iated Hosp ital of Chongq ing Medical Unive rs ity, Chongq ing 400016, China ） Abstract: 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 dia betes 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 spe cific for TCF7L2 to verify the binding of TCF7L2 to IDE promoter. Results indicated that IDE gene specif ic fragments were detected in the DNA fragments immunoprecip