基础医学与临床 ›› 2018, Vol. 38 ›› Issue (3): 355-360.

• 研究论文 • 上一篇    下一篇

转录因子2过表达改善人肝癌细胞HepG2的胰岛素抵抗

权晓娟1,胡艳粉2,梁春联2,郑华东2   

  1. 1. 西安交通大学医学院第二附属医院
    2. 西安交通大学第二附属医院
  • 收稿日期:2017-03-13 修回日期:2017-05-24 出版日期:2018-03-05 发布日期:2018-02-27
  • 通讯作者: 权晓娟 E-mail:xiaojuanquan@163.com
  • 基金资助:
    陕西省社会发展科技攻关项目;西安市2015年社会发展引导计划-医学研究项目

Overexpression of TCF2 ameliorates insulin resistance in human hepatocellular carcinoma cells HepG2

  • Received:2017-03-13 Revised:2017-05-24 Online:2018-03-05 Published:2018-02-27

摘要: 目的 研究转录因子2(TCF2)过表达对HepG2细胞胰岛素抵抗的影响。方法 用高浓度胰岛素(1×10-8 mol/L)诱导处理人肝癌HepG2细胞24 h建立人肝癌HepG2细胞胰岛素抵抗模型,分为空白组、胰岛素抵抗模型组(IR组)、IR+空载体组和IR+TCF2过表达组;RT-qPCR和Western blot检测TCF2的表达;葡萄糖氧化酶法检测培养液中的葡萄糖浓度;蒽酮法检测糖原合成量;MTT法检测细胞存活率;比色法检测己糖激酶和丙酮酸激酶的活性;Western blot检测人胰岛素受体底物(IRS-1)和葡萄糖转运蛋白4(GLUT4)的表达。结果 与空白组比较,IR组细胞葡萄糖消耗量显著降低(P<0.05),表明模型建立成功。IR组细胞TCF2 mRNA和蛋白表达显著降低(P<0.05)。与IR组相比,TCF2过表达可显著增加胰岛素抵抗HepG2细胞的葡萄糖消耗量、肝糖原合成量、己糖激酶活性和丙酮酸激酶活性,增加IRS-1和GLUT4蛋白表达(P<0.05)。结论 过表达TCF2可改善人肝癌HepG2细胞的胰岛素抵抗。

关键词: TCF2, 胰岛素抵抗, HepG2细胞

Abstract: Objective To investigate the effects of TCF2 overexpression on insulin resistance in HepG2 cells. Methods HepG2 cells were treated with high concentration of insulin (1×10-8 mol/L) for 24 hours to induce insulin resistance (IR). Cells were divided into four groups: control group, IR group, IR+vector group and IR+TCF2 overexpression group. RT-qPCR and Western blot were performed to detect the expression of TCF2. Glucose consumption and glycogen synthesis were assayed by glucose oxidase method and anthrone method, respectively. Cell viability was evaluated by MTT assay. The activities of hexokinase and pyruvate kinase were detected by colorimetry. The protein level of IRS-1 and GLUT4 was detected by Western blot. Results Compared with control group, the decreased glucose consumption was observed in IR group (P<0.05), indicating that insulin-resistance model was established successfully. The mRNA and protein expression of TCF2 was remarkably down-regulated in IR group compared with control group. Compared with IR group, overexpression of TCF2 significantly improved glucose consumption, liver glycogen content, and the activities of hexokinase and pyruvate kinase (P<0.05). Moreover, TCF2 overexpression up-regulated the protein expression of IRS-1 and GLUT4 (P<0.05). Conclusion TCF2 overexpression ameliorates insulin resistance of HepG2 cells.

Key words: TCF2, insulin resistance, HepG2 cells