SiRNA沉默HBx表达对HepG2.2.15细胞增殖和凋亡的影响

The effects of silencing of HBx gene expression by siRNA on the prolifer ation and apoptosis of HepG 2. 2. 15 cells

目的观察沉默HB x基因表达对肝癌细胞HepG2.2.15增殖和凋亡的影响。方法用脂质体转染法将pSIHBV/X转染肝癌细胞HepG2.2.15,RT-PCR法检测HBx mRNA表达;ELISA法检测细胞上清液中HBsAb和HBeAg的表达情况;MTT法检测对细胞增殖的影响;AnnexinV-FITC/PI双染法检测细胞凋亡情况。结果 pSIHBV/X质粒转染HepG2.2.15细胞后HBx mRNA表达较对照组明显降低(P<0.01);G2.2.15细胞上清液中HBsAg和HBeAg的水平下降,与对照组比较差异有统计学意义(P<0.01);MTT法检测结果显示转染pSIHBV/X质粒后HepG2.2.15细胞的增殖受到抑制,AnnexinV-FITC/PI双染法检测结果显示HBx siRNA可促进细胞凋亡,与对照组比较差异有统计学意义(P<0.01)。结论靶向HBx基因的干扰质粒能降低HepG2.2.15细胞中HBsAg和HBeAg的水平、抑制HepG2.2.15细胞增殖,促进HepG2.2.15细胞凋亡。

Objective Observe the effects of silencing HBx gene expression on the proliferation and apoptosis of HepG2.2.15 cells.Methods pSIHBV / X siRNA interference vectors were transfected into HepG2.2.15 cells.The HBx mRNA expression was measured using RT-PCR.The levels of HBsAg and HBeAg in cell supernatant were determined by ELISA.The cell proliferation was evaluated by MTT.The cell apoptosis was detected by Annexin V-FITC / PI double staining method.Results After transfection of the pSIHBV / X siRNA vectors,HBx mRNA expression was inhibited compared with control group（P 0.01）;The levels of both HBsAg and HBeAg in HepG2.2.15 cell supernatants were significantly decreased after the transfection of pSIHBV / X,compared with the control group（P 0.01）.The cell proliferation was inhibited and the cell apoptosis was promoted after the transfection of pSIHBV / X,compared with the control group（P 0.01）.Conclusion Our results display that HBx gene RNA interference resulted in the reduction of the levels of HBsAg and HBeAg,the inhibition of cell proliferation and the increase of cell apoptosis in HepG2.2.15 cells.