南方医科大学学报

南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (12): 1700-.

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miR-205通过下调ZEB1和ZEB2表达抑制肾小管上皮细胞转分化

曹罗元，杨菁，富显果，林应华，林峰，黄宝英   

  • 出版日期:2016-12-20 发布日期:2016-12-20

The MicroRNA miR-205 inhibits epithelial-messenchymal transition in HK-2 cells by down-regulating ZEB1 and ZEB2 expressions

  • Online:2016-12-20 Published:2016-12-20

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摘要： 目的探讨miR-205在肾小管上皮细胞转分化的作用机制。方法将miR-205 mimics和scrambled control分别转染HK-2 转分化细胞株，采用real-time qPCR检测miR-205 和ZEB1、E-cadherin、α-SMA mRNA的表达水平；运用Western blotting 检测 ZEB1、ZEB2、E-cadherin、α-SMA的表达水平。通过细胞免疫组化检测β-catenin的异位表达情况和E-cadherin的表达情况。结 果miR-205 mimics 转染HK-2转分化细胞株后，ZEB1和ZEB2的表达水平较高糖组得到大幅下调（P<0.01），而E-cadherin的 表达水平较高糖组显著提高（P<0.01），同时间质细胞特征分子α-SMA的表达水平显著降低（P<0.01）。miR-205 mimics 也显著 抑制了HK-2 转分化过程中β-catenin 异位表达，在维持上皮细胞的形态方面也起着重要的作用。结论miR-205 可通过下调 ZEB1和ZEB2的表达，抑制了肾小管上皮间质转分化进程。

Abstract: Objective To explore the role of miR-205 in regulating epithelial-messenchymal transition (EMT) in proximal tubular cell line HK-2 cells and the underlying mechanism. Methods HK-2 cells transfected with miR-205 mimics or a scrambled control sequence were examined for miR-205 expressions and mRNA levels of ZEB1, E-cadherin, and α-SMA using real-time qPCR; the protein levels of ZEB1, ZEB2, E-cadherin, and α-SMA were detected with Western blotting. Immunohistochemistry was performed to examine the ectopic expression of β-catenin and E-cadherin expression in the cells. Results The expression levels of ZEB1 and ZEB2 decreased significantly (P<0.01) while E-cadherin expression was up-regulated (P<0.01) in cells transfected with miR-205 mimics. Transfection with miR-205 mimics also markedly down-regulated the expression of α-SMA (P<0.01), a marker of mesenchymal cells that play an important role in EMT of HK-2 cells. The ectopic expression of β-catenin was inhibited by miR-205 mimics in HK-2 cells. Conclusion miR-205 inhibits EMT in HK-2 cells by down-regulating the expression levels of ZEB1 and ZEB2.