干扰NBS1表达抑制肝癌HepG2细胞增殖并促进凋亡

目的:探讨干扰NBS1基因表达对肝癌细胞HepG2增殖及凋亡的影响。方法:应用Lipofectamine 2000将NBS1特异性小干扰RNA（siNBS1）转染至肝癌细胞HepG2中，以空载脂质体作为对照组。采用RT-PCR及Western blot法检测NBS1基因表达情况，采用CCK-8法检测转染质粒后siNBS1对HepG2细胞增殖能力的影响，采用流式细胞术检测siNBS1对HepG2细胞凋亡的影响。结果:不同浓度siNBS1（10 nmol/L、20 nmol/L、50 nmol/L）均能抑制HepG2细胞中的NBS1基因在mRNA水平上的表达（P＜0.05或P＜0.01）。最高浓度siNBS1（50 nmol/L）能抑制NSB1蛋白产物的表达水平（P＜0.05）。CCK-8实验显示，转染不同浓度siNBS1的HepG2细胞与对照组相比，细胞活力明显减弱（P＜0.05或P＜0.01），转染48 h后增殖率下降最为显著；流式细胞检测结果显示，细胞凋亡的比率在转染后明显升高（P＜0.05或P＜0.01），并随浓度增加而更为显著。结论:干扰NBS1基因的表达可以明显抑制HepG2细胞的增殖，并促进癌细胞凋亡。

Interfering NBS1 expression inhibits proliferation and promotes apoptosis of liver cancer HepG2 cells

Objective:To investigate the effect of NBS1 knockdown on proliferation and apoptosis of HepG2 cells.Methods:NBS1 specific small interference RNA(siNBS1) was transfected into HepG2 cells,performed using Lipofectamine 2000,with empty liposome as control group.The expression of NBS1 was detected by RT-PCR and Western blot.The HepG2 cell proliferation was detected by CCK-8 method,and cell apoptosis was analyzed by using FCM.Results:Different concentrations of siNBS1(10 nmol/L,20 nmol/L,50 nmol/L) could inhibit the expression of NBS1 gene at mRNA level in HepG2 cells(P＜0.05 or P＜0.01).The highest concentration of siNBS1(50 nmol/L) could significantly inhibit the expression of NSB1 protein.CCK-8 assay demonstrated that the proliferation of HepG2 cells in siNBS1 group transfected with different concentrations of siNBS1 was lower than that of the control group(P＜0.05 or P＜0.01),and the proliferation rate decreased most significantly after 48 h of transfection.FCM revealed that the apoptosis rate increased significantly after transfection(P＜0.05 or P＜0.01),and was related to the concentration.Conclusion:Inhibition of NBS1 gene expression can significantly inhibit the proliferation and promote apoptosis of HepG2 cells.