RNA干扰沉默mTOR基因对肝癌HepG2细胞增殖和凋亡的影响

Effect of mTOR gene inhibition by siRNA on proliferation and apoptosis of human hepatoma HepG2 cells

目的:探讨RNA干扰技术对人肝癌HepG2细胞mTOR的表达及细胞增殖与凋亡的影响。方法:体外合成mTOR siRNA,并转染HepG2细胞,同时设无义对照组(转染无义siRNA)、空白对照组(转染空脂质体)及正常对照组(不转染)。采用western blot法检测各组HepG2细胞的mTOR蛋白表达;应用MTT法检测细胞增殖;应用TUNEL法检测细胞凋亡,计算凋亡指数。结果:mTOR siRNA转染组HepG2细胞mTOR蛋白的表达显著低于各对照组(P<0.05);mTOR siRNA转染组HepG2细胞增殖低于各对照组(P<0.05);mTOR siRNA转染组细胞凋亡指数高于各对照组(P<0.05)。结论:mTOR siRNA对HepG2细胞中mTOR蛋白的表达有明显的抑制作用,且能显著抑制HepG2细胞的增殖并促进其凋亡。

Objective：To study the effects of mTOR siRNA on the expression of mTOR gene,cell proliferation and apoptosis of human hepatoma HepG2 cell.Methods：The oligonucleotide templates of mTOR siRNA were synthesized and used to transfected HepG2 cells.Three control groups were established accordingly by transfection of siRNA with insignificant order,liposome alone or no treatment,respectively.The protein levels of mTOR were measured by western blot method,cell proliferation and apoptosis were detected with MTT and TUNEL method,respectively.Results：The expression of mTOR protein in the experiment group that was transfected with mTOR siRNA was lower than those of the control groups（P〈0.05）.Proliferation of HepG2 cells transfected with mTOR siRNA were lower than those of the control groups（P〈0.05）,while the apoptosis index were higher（P〈0.05）.Conclusion：mTOR siRNA shows significant inhibition on the expression of mTOR protein and proliferation of HepG2 cells,as well as promotion of its apoptosis.