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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 effect of down-regulation of c-Met expression on invasion and growth of hepatocellular carcinoma (HCC) cells. Methods: The c-Met gene of HCC cell line Bel-7402 and HepG2 was silenced by siRNA. The cell counting kit (CCK-8) was used to detect the cell proliferation and growth. The changes of cell cycle were detected. Transwell chamber assay was used to detect cell invasion. Western blot was used to detect the expression of Ras, Raf, p-Erk, cyclin D1, β-catenin and Slug. Results: siRNA technology successfully down-regulated the c-Met gene expression of Bel-7402 and HepG2. After down-regulating the expression of c-Met, the proliferation activity of HCC cells decreased, and the G1 phase arrest obviously occurred in the early stage of DNA synthesis, the invasion ability weakened and the expressions of Raf, p-Erk, cyclin D1, β-catenin and Slug decreased. Conclusion: Down-regulation of c-Met gene expression in HCC cells can inhibit cell proliferation and growth, arrest the cell cycle and weaken cell invasion. The mechanism may be related to the interference of Ras / Raf / ERK, Wnt pathway and EMT process.