siRNA抑制NFBD1基因表达对人肝癌HepG2细胞凋亡的影响

Effects of siRNA-mediated NFBD1 silencing on cellular apoptosis in HepG2 cells

目的:研究siRNA抑制NFBD1基因表达对人肝癌HepG2细胞凋亡的作用原理及caspase途径在此过程中的作用。方法:应用流式细胞法及RT-PCR检测沉默NFBD1对人肝癌HepG2细胞凋亡及凋亡基因组表达水平的影响。MTT法检测NFBD1 siRNA对HepG2细胞的抑制作用。结果:转染48 h后,转染组与阴性对照组和空白组相比,NFBD1mRNA表达明显减弱,转染组NFBD1 mRNA较空白组相对下降74.75%;HepG2细胞组中促凋亡基因Bc1-xl表达减少,Bid及caspase-3表达量增加(P<0.05)。MTT结果显示NFBD1 siRNA能明显抑制HepG2细胞增殖,抑制率达到92.131±0.893%。流式法检测结果显示转染组细胞凋亡明显高于阴性对照组和空白组。结论:NFBD1可通过改变Bc1-xl/Bid的表达比,激活caspase-3来诱导HepG2细胞凋亡。

Objective：To investigate the role and rationale of NFBD1 in cell apoptosis,and to examine the role of caspase pathway in the apoptosis process,by using the specific siRNA duplexes to inhibit NFBD1 expression in HepG2 cells.Methods： Using flow cytometry（FCM） and RT-PCR to check the expression of apoptosis-associated genes and effect of cell apoptosis in HepG2 cells with NFBD1 siRNA.The inhibitory actions of HepG2 cell growth with NFBD1 siRNA are measured by MTT assay.Results： The expression of NFBD1 mRNA of transfection group was lower than that of negative control group and blank group at 48 h after transfection.The expression of NFBD1 mRNA of transfection group fell 74.75 percent.when compared to that of blank group.The expression of antiapoptotic mitochondrial gene bcl—xl decreased,and proapoptotic gene bid and caspase-3 increased in transfection group（P 0.05）.MTT assay suggests that NFBD1 siRNA can inhibit the growth of HepG2 cell seriously,and the inhibiting rate of it is 92.131±0.893%.The result of flow cytometry（FCM）shows that apoptosis rate of transfection group was higher than that of negative control group and blank group obviously.Conclusion：NFBD1 may induce HepG2 cell apoptosis through alteration of the ratio of Bc1-xl/Bid and activation of caspase-3.