ASPP2抑制HBx引起细胞自噬的实验研究

目的 观察ASPP2对HBx引起的细胞自噬的改变.方法 运用Hbx转染HepG 2细胞,ASPP2过表达与对照情况下,采用免疫印迹技术检测内源性自噬相关蛋白LC3Ⅱ/LC3 Ⅰ和P62的表达,采用免疫荧光方法检测LC3荧光颗粒表达水平,采用RT-PCR技术检测自噬相关基因Beclin-1的mRNA转录水平.结果 免疫印迹技术显示转染HBx质粒比转染空载组的LC3Ⅱ量增多,但是在加入ASPP2腺病毒组比空载组的LC3Ⅱ表达量减少.免疫荧光显示转染HBx质粒比转染空载组的LC3荧光颗粒多,但是在共转染ASPP2质粒组比空载组的LC3荧光颗粒减少.RT-PCR方法显示转染HBX质粒比转染空载组的表达增多,但是在加入ASPP2腺病毒组比空载组表达量减少.结论 ASPP2可以抑制HBx引起的细胞自噬.

The experiment research about using ASPP2 to inhibit the autophagy caused by HBx

Objective To investigate the effect of ASPP2 on autophagy caused by HBx.Methods In condion of over-expession of ASPP2 and control,HepG 2 cells were transfected with HBx plasmid,endogenous LC3 protein were detected by western blotting.LC3 punctas in HBV infected cells were counted by immunofluorescence.mRNA levels of autophagy related gene Beclin-1 were tested by realtime-PCR (RT-PCR).Results Immunoblotting showed that the amount of LC3 was increased in the transfected HBx plasmid compared with the transfected no-load group,but the expression of LC3 Ⅱ was lower in the Aspp2 adenovirus group than in the empty group.Immunofluorescence showed that the transfected HBx plasmid was more abundant than the LC3 fluorescent particles in the transfected no-load group,but decreased in the co-transfected ASPP2 plasmid group than the empty group.RT-PCR showed that the expression of HBx was higher than that of transfection group (P ＜ 0.05).Conclusion ASPP2 could inhibit the autophagy caused by HBx.