乙型肝炎病毒HBx蛋白抑制TTRAP启动子活性的体外研究  

Repression of TRAP Promoter by HBx Activity in vitro

作　　者：张小花[1] 张磊[1] 田园园[1] 王丽英[1] 黄爱龙[1] 汤华[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《生物技术通报》2010年第2期135-139,172,共6页Biotechnology Bulletin

基　　金：国家自然科学基金(30771924)

摘　　要：旨在探讨体外培养条件下HBx蛋白对TTRAP(TRAF and TNF receptor-associated protein)基因转录水平表达的影响。用RT-PCR及Real-time PCR检测TTRAP在HepG2细胞和HepG2.2.15细胞中的表达;构建TTRAP启动子虫荧光素酶报告质粒;分别与HBV、HBs、HBp、HBc、HBx表达质粒共转染HepG2细胞,比较虫荧光素酶活性。RT-PCR和Real-time PCR结果显示,TTRAP在HepG2.2.15细胞中的表达量分别是其在HepG2细胞中表达量的44.9%和27.8%(P<0.05)。TTRAP启动子虫荧光素酶报告质粒与HBV表达质粒共转染组的相对荧光素酶活性,与对照组相比下降了43.8%。转染HBx表达质粒组的相对荧光素酶活性与其对照组相比下降了35%,而转染HBc、HBs及HBp表达质粒组对相对荧光素酶活性没有影响。因此证实HBx蛋白能抑制TTRAP启动子活性。It was to study the effect of HBx on TTRAP transcriptional level expression in vitro, and explore the relationship between HBx and transcriptional level expression of TTRAP. RT-PCR and Real-time PCR were performed to check TTRAP expression in HepG2 and HepG2.2.15 cell lines. TTRAP promoter lueiferase reporter plasmid,pGL3-Basie-TTRAP-P,was constructed. HepG2 cells were transiently cotransfected with the same amount of pGL3-Basie-TTRAP-P and HBV or HBx, HBs, HBe, HBp expression plasmids, respeetively. Then the lueiferase activity was detected. RT-PCR and Real-time PCR results showed that the amounts of TTRAP expression in HepG2.2. 15 eells are 44.9% and 27.8% , lower than that in HepG2 cells, respectively. The relative luciferase activity in HepG2 cells,which were transiently eotransfeeted with pGL3-Basie-TTRAP-P and HBV expression plasmid,reduced 43.8% lower than that in control group. The relative lueiferase activity in HepG2 cells transfected with HBx expression plasmid reduced 35% comparison to the control group. However, HBs, HBe and HBp expression plasmids had no effect on lueiferase activity. We found that HBx can repress TTRAP promoter activity in vitro.

关 键 词：HBX蛋白 抑制 TTRAP 启动子活性 HEPG2.2.15细胞 HEPG2细胞 

分 类 号：R512.62[医药卫生—内科学]