Yang Yida, Zheng Lin, Lv Guocai, Jin Xi, Sheng Jifang
State Key Laboratory for Infectious Diseases Diagnosis and Treatment, Department of Infectious Diseases, The First Affiliated Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang, PR China.
Intervirology. 2007;50(5):323-7. doi: 10.1159/000106463. Epub 2007 Jul 26.
To investigate the effect of HBV X protein (HBX) on TNF-alpha-related apoptosis-induced ligand (TRAIL) expression in HepG2 cells, and observe death of primary hepatocytes induced by HBX-transfected HepG2 cells.
Western blot was used to detect the TRAIL expression in HepG2 cells transfected with mammalian expression plasmid pSG5UTPL-HBX. The reverse transcription-PCR was used to observe TRAIL mRNA transcription stimulated by HBX protein, and chromium release assay was used to detect death of primary hepatocyte induced by HBX-transfected HepG2 cells.
HBX could increase TRAIL expression and mRNA transcription in HepG2 cells in a dose-dependent manner. The C-terminal truncated version of HBX (HBXD1) is responsible for inducing TRAIL expression in HepG2 cells. Chromium release assay results showed that HBX-transfected HepG2 cells kill primary human hepatocytes by a TRAIL-mediated mechanism. Neutralizing anti-TRAIL inhibits the HepG2 killing.
HBX protein increases TRAIL expression in HepG2 cells which induced death of primary hepatocytes. HBX protein may play an important role in mechanisms of hepatic cell death and hepatic inflammation caused by HBV infection.
研究乙肝病毒X蛋白（HBX）对HepG2细胞中肿瘤坏死因子-α相关凋亡诱导配体（TRAIL）表达的影响，并观察HBX转染的HepG2细胞诱导的原代肝细胞死亡情况。
采用蛋白质免疫印迹法检测用哺乳动物表达质粒pSG5UTPL-HBX转染的HepG2细胞中TRAIL的表达。采用逆转录-聚合酶链反应观察HBX蛋白刺激的TRAIL mRNA转录情况，并用铬释放试验检测HBX转染的HepG2细胞诱导的原代肝细胞死亡情况。
HBX可呈剂量依赖性增加HepG2细胞中TRAIL的表达和mRNA转录。HBX的C末端截短型（HBXD1）负责诱导HepG2细胞中TRAIL的表达。铬释放试验结果表明，HBX转染的HepG2细胞通过TRAIL介导的机制杀伤原代人肝细胞。中和抗TRAIL可抑制HepG2细胞的杀伤作用。
HBX蛋白增加HepG2细胞中TRAIL的表达，从而诱导原代肝细胞死亡。HBX蛋白可能在乙肝病毒感染引起的肝细胞死亡和肝脏炎症机制中起重要作用。
