下调c–Met表达对肝癌细胞侵袭生长能力的影响

Effects of Down-regulation of c-Met Expression on Invasion and Growth of Hepatocarcinoma Cells

目的:探讨下调c–Met表达对肝癌(HCC)细胞侵袭生长能力的影响。方法:采用小干扰RNA(siRNA)技术沉默HCC细胞株Bel–7402和HepG2的c–Met基因,应用细胞计数试剂盒(CCK–8)方法:克隆形成实验检测细胞增殖和生长,流式细胞仪检测细胞周期的变化,Transwell小室实验检测细胞侵袭能力,蛋白质印迹法(Western blot)方法检测相关信号通路蛋白Ras、Raf、p–Erk、cyclin D1、β–catenin和Slug的表达。结果:siRNA技术能成功下调Bel–7402和HepG2的c–Met基因表达。下调c–Met表达后,HCC细胞增殖活性下降,发生明显DNA合成前期G1期停滞,侵袭能力减弱,Raf、p–Erk、cyclin D1、β–catenin和Slug蛋白表达下降。结论:下调HCC细胞c–Met基因的表达能够抑制细胞增殖和生长,阻滞细胞周期,减弱细胞侵袭能力,其机制可能与干扰Ras/Raf/ERK、Wnt通路和EMT过程有关。

Objective To investigate the effects of c–Met down–regulation on HCC cells growth and invasion. Methods RNA interference technique was applied to knockdown c–Met expression in HCC cell lines,including Bel–7402 and HepG2. HCC cell proliferation was evaluated by CCK–8 method and colony formation assay. Cell cycle was detected by flow cytometry. Cell invasion was analyzed using Transwell assay with Matrigel. Western blot analysis was applied to study the expression of Ras, Raf, p–Erk, cyclin D1, β–catenin and Slug. Results C–Met expression in HCC cells was down–regulated by si RNA technique. Inhibition of c–Met expression significantly suppressed HCC cells proliferation,growth and invasive capacity, and arrested cell cycle in the G1 phase,as well as repressed some important proteins expression,including Raf,p–Erk,cyclin D1,β–catenin and Slug. Conclusions Down–regulation of c–Met expression obviously inhibits HCC cells proliferation, growth, cell cycle and invasive capacity, which probably related to suppression of Ras/Raf/ERK pathway, Wnt pathway and EMT process.