MicroRNA-100调节mTOR表达对肝癌细胞侵袭转移及上皮间质转化的影响及机制研究

MicroRNA- 100 regulates Hep G2 cell migration,invasion and EMT process by regulating mTOR gene expression

目的:探讨microRNA-100调节m TOR表达对肝癌细胞侵袭转移及上皮间质转化的影响并研究其可能的分子机制。方法:通过脂质体介导将microRNA-100模拟物及阴性对照转染Hep G2细胞,将细胞分为转染试剂对照组(mock)、阴性对照组(control)和microRNA-100 mimic(miRNA-100 mimic)组。采用realtime PCR检测转染后细胞中miRNA-100的表达水平,Western blot检测m TOR的表达变化,细胞划痕实验检测细胞的迁移能力,Tranwell检测细胞的侵袭能力,Western blot检测细胞中MMP-2、MMP-9、N-cadherin、Vimentin、α-SMA和E-cadherin的表达。结果:过表达miRNA-100通过抑制m TOR的表达抑制肝癌细胞的迁移及侵袭能力,并通过下调肝癌细胞中MMP-2、MMP-9、N-cadherin、Vimentin和α-SMA的表达,上调E-cadherin的表达,阻抑上皮细胞-间质转化(epithelial-mesenchymal transition,EMT)过程的发生。结论:MicroRNA-100可能通过降低m TOR的表达抑制肝癌细胞的转移侵袭能力,并抑制上皮间质转化过程的发生。

Objective：To investigate the effect of microRNA-100 on the migration,invasion and epithelial-mes-enchymal transition（ EMT）process of human hepatocarcinoma HepG2 cells and the possible mechanism. Methods：Synthetic miRNA-100 mimic and its negative control were transfected into HepG2 cells by liposome method. We di-vided cells into three groups：mock group,control group and miRNA-100 mimic group. After transfection,the ex-pression of mTOR at mRNA level was detected by quantitative real-time PCR. The migration of HepG2 cells was de-tected by wound scratch assay,and the invasion ability changes were analyzed in transwell. Expressions of MMP-2, MMP-9,N-cadherin,Vimentin,α-SMA and E-cadherin were analyzed by Western blot. Results：MiRNA-100 can inhibit the migration and invasion ability via down -regulate the mTOR expression. The Western blot results showed that miRNA-100 could down-regulate the expression of MMP-2,MMP-9,N-cadherin,Vimentin and α- SMA and up - regulated the expression of E - cadherin,and inhibit the progress of EMT. Conclusion：MiRNA -100 can suppress the migration and invasion process and EMT process,it is may be achieved by down - regulating mTOR gene expression.