沉默SEPT9基因对肝癌HepG2细胞增殖及凋亡的影响

Effects of SEPT9 Gene Silencing on Cell Proliferation and Apoptosis of HepG2 Hepatoma Cells

目的探讨SEPT9基因与肝癌的关系,进一步揭示SEPT9基因在肝癌发生发展过程中的作用。方法将构建的针对SEPT9基因的shRNA表达载体通过脂质体LipofectamineTM2000转染人肝癌HepG2细胞,荧光显微镜下观察细胞的转染效率,RT-PCR、Western blot法分别检测SEPT9 mRNA和蛋白质的表达抑制情况,CCK-8检测细胞的生长抑制情况,流式细胞术检测细胞凋亡情况。结果荧光显微镜观察细胞的转染效率达到70%以上;SEPT9 mRNA及蛋白表达抑制率均受到明显抑制。CCK-8法检测结果显示,实验组HepG2细胞的生长受到明显抑制;流式细胞术检测结果显示实验组HepG2细胞凋亡明显(P<0.05)。结论抑制SEPT9基因的表达可有效抑制HepG2细胞的增殖,并对细胞凋亡有明显的促进作用。

Objective To explore the relationship between SEPT9 gene and hepatocellular carcinoma,and to reveal the role of SEPT9 in tumorigenesis and development of hepatoma.Methods The expression vector containing SEPT9 gene was transfected into the human hepatoma cell line HepG2 with LipofectamineTM2000.The transfection efficacy was studied under fluorescence microscopy,and the mRNA and protein levels of SEPT9 were detected with reverse transcript polymerase chain reaction（RT-PCR） and Western blot assay.CCK-8 method was used to detect cell proliferation,and FCM was served to detect the cell apoptosis.Results The transfection efficiency was up to 70%,and expression of mRNA and protein of SEPT9 were suppressed significantly.And the result of CCK-8 assay showed significantly inhibition of in the experiment group,and FCM assay showed that the apoptotic rate in the experiment group increased significantly comparing to the control group（P0.05）.Conclusion Suppression of the expression of SEPT9 gene can effectively inhibit the proliferation of HepG2 cell,and promote apoptosis significantly.