shRNA干扰GRP78基因对肝癌细胞株SMMC-7721增殖和凋亡的影响

shRNA-GRP78 inhibited proliferation and induced apoptosis of liver cancer SMMC-7721 cells

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-antiGRP78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 cells proliferation and induced their apoptosis.