shRNA干扰GRP78基因对肝癌细胞株SMMC-7721增殖和凋亡的影响

摘要

目的:研究shRNA干扰GRP78基因对肝癌细胞株SMMC-7721增殖和凋亡的影响.方法:将GRP78特异性shRNA质粒载体Pla-anti-GRP78转染SMMC-7721,采用流式细胞术(FCM)检测转染效率、分析细胞周期分布和凋亡,从蛋白和mRNA水平检测干扰GRP78效果,MTT法检测干扰GRP78对SMMC-7721增殖的影响.结果:Pla-anti-GRP78转染SMMC-7721细胞48h后,GRP78表达下降,空白对照组GRP78 mRNA的表达量为1,无关对照组组为0.95,而Pla-anti-GRP78组为0.25(P ＜0.05);转染Pla-anti-GRP78的SMMC-7721细胞G2期阻滞(55.2％,P＜0.05);空白对照组凋亡率为6.6％,无关对照组为8.1％,而Pla-anti-GRP78组高达58.2％ (P ＜0.05).结论:shRNA干扰GRP78表达抑制SMMC-7721的增殖,并诱导其凋亡.%Objective:To evaluate the effect of GRP78 special shRNA on proliferation and apoptosis of human liver cancer SMMC-7721 cells.Methods:The expression vector of Pla-anti-GRP78 was constructed and transfected into SMMC-7721 by Lipofectin.The protein and mRNA expression of GRP78 was detected by Western blotting and real-time PCR.The changes of cell cycle after transfection were examined by flow cytometry and MTT assay,respectively.Results:In SMMC-7721 cells,the protein and mRNA expression levels of GRP78 were significantly decreased after transfection,and reduction of proliferation was related to an increase in the fraction of G2 phase.Conclusion:The GRP78 special siRNA silenced GRP78,decreased SMMC-7721 ceils proliferation and induced their apoptosis.