siRNA靶向沉默Rictor基因表达对肝癌细胞增殖、迁移和侵袭的影响

目的 探讨靶向沉默Rictor表达对肝癌细胞增殖、迁移和增殖的影响。方法 采用LipofectamineTM2000向SMMC-7721和Hep3B细胞转染成功构建靶向沉默Rictor表达的siRNA载体片段（沉默组）或无义siRNA片段（对照组）,转染48 h后采用QPCR检测Rictor mRNA水平,以Western blotting检测Rictor、p-Akt、细胞周期蛋白和EMT相关蛋白的表达情况。MTT实验、EdU增殖实验、Transwell实验和Boyden实验分别检测肝癌细胞的增殖、迁移和增殖能力。结果 转染48 h后,沉默组细胞的Rictor mRNA蛋白水平大部分都低于对照组细胞;沉默组细胞的增殖、迁移、侵袭能力均低于对照组细胞,差异有统计学意义（P<0.01）。MTT实验及EdU增殖实验表明,两组肝癌细胞在沉默Rictor表达后增殖减慢、S期细胞比例明显减少（P均<0.01）;Transwell实验和Boyden实验表明,两组肝癌细胞在沉默Rictor表达后穿膜细胞数显著减少（P均<0.01）。结论 Rictor基因在肝癌细胞中起到癌基因的作用,沉默Rictor的表达抑制肝癌细胞的增殖、迁移和侵袭的过程,可能与下调p-Akt、CCND1和N-cadherin、ZEB1、Vimentin蛋白表达,上调E-cadherin,P21和P27蛋白表达有关。

Effects of siRNA silencing Rictor gene expression on proliferation,migration and invasion of hepatocellular carcinoma cells

Objective To investigate the effect of siRNA targeted silencing Rictor expression on the proliferation, migration and proliferation in hepatocellular carcinoma cells. Methods LipofectamineTM2000 was used to transfect SMMC-7721 and Hep3B cells to construct siRNA vector fragments (silenced group) or sense-free siRNA fragments (control group) targeting Rictor expression. After transfection for 48 h, Rictor mRNA level was detected by QPCR and Rictor, p-Akt, cyclin and EMT-related proteins were detected by Western blotting. MTT assay, EdUincorporation assay, Transwell assay and Boyden assay were used to detect the proliferation, migration and proliferation of HCC cells, respectively. Results After transfection for 48 h, most of the Rictor mRNA level of the silenced group was lower than that of the control group. The ability of cell proliferation, migration and invasion in the silenced group was weakened when compared to that in the control group (P<0.01). In MTT assay and EdU incorporation assay, knockdown of Rictor expression inhibited proliferation in two hepatocellular carcinoma cell lines, while in Transwell assay and Boyden assay, knockdown of Rictor expression inhibited migration and invasion in both cell lines (P<0.01). Conclusion Rictor gene plays an oncogene role in hepatocellular carcinoma cells. Silencing Rictor expression inhibits the proliferation, migration and invasion of hepatocellular carcinoma cells, which may be related to down-regulating the expression of p-Akt, CCND1 and N-cadherin, ZEB1 and Vimentin, and up-regulating the expression of E-cadherin, P21 and P27.