HBV转染对TWEAK诱导凋亡的影响及IFN-γ对凋亡的调节

Effect of HBV transfection on apoptosis induced by TNF-like weak inducer of apoptosis and the regulatory effect of IFN-γ on apoptosis

目的:探讨新型凋亡分子TNF-like weak inducer of apoptosis(TWEAK)对乙型肝炎病毒(HBV)转染的人肝癌细胞系的凋亡诱导作用及γ-干扰素(IFN-γ)对该凋亡过程的影响。方法:以稳定转染了pCD-NA3/1.1HBV的肝癌细胞系BEL-7402-pCDNA3/1.1HBV为模型,并以稳定转染空载体的肝癌细胞BEL-7402/pcDNA3作为对照,经CCK-8法检测HBV转染前后TWEAK对细胞生长的抑制效应,terminal transferase dUTP nick end labeling(TUNEL)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)活性检测试剂盒检测HBV转染前后TWEAK诱导肝癌细胞的凋亡情况及IFN-γ对TWEAK诱导细胞凋亡的调节作用。结果:HBV转染促进TWEAK诱导的细胞凋亡,IFN-γ可以增强HBV转染细胞对TWEAK诱导凋亡的敏感性。结论:机体感染HBV后,TWEAK可能参与HBV感染肝细胞的清除过程;IFN-γ在调节TWEAK诱导的病毒感染细胞凋亡中发挥重要作用。

AIM： To explore the effect of HBV transfection on apoptosis induced by TWEAK and the regulatory effect of IFN - γ on the apoptosis process. METHODS ： BEL - 7402 - pCDNA3/1.1HBV hepatoma cell line was used as cell model in this experiment, in which BEL -7402 was stably transfected with pCDNA3/1. 1HBV and its corresponding empty vector BEL - 7402 - pCDNA3 was used as control. The effect of HBV transfection on TNF - like weak inducer of apoptosis（TWEAK） induced hepatoma cell apoptosis was determined by CCK -8. The apoptosis before and after HBV trans- fection induced by TWEAK and the effect of IFN -γ on apoptosis induced by TWEAK in HBV transfected hepatoma cells were detected by TUNEL and caspase 3 activity detection kit. RESULTS： BEL -7402 -pCDNA3/1. 1HBV and BEL - 7402 - pCDNA3 hepatoma cell line were successfully established and identified. Cell viability of hepatoma cells was increased after HBV transfection. The apoptosis rate was much higher after HBV transfection compared to the empty vector control and empty cell control. The apoptosis rate induced by TWEAK was much higher in BEL - 7402 - pCDNA3/1. 1HBV ceils when exposed to IFN -γ. CONCLUSION： After HBV infection, TWEAK might be involved in the clearance of HBV infected hepatocyte by apoptosis and inflammatory factor such as IFN - γ.