HBV通过增强DNAJB4启动子活性调节其在HepG2．2．15细胞中的表达  

HBV Regulates DNAJB4 Expression in HepG2. 2. 15 Cells through Enhancing its Promoter Activity

作　　者：张小花[1] 万涛[1] 张磊[1] 田园园[1] 黄婷婷[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,重庆市400016

出　　处：《医学分子生物学杂志》2009年第5期391-396,共6页Journal of Medical Molecular Biology

基　　金：国家自然科学基金（No.30771924）

摘　　要：目的初步探讨HBV与DNAJB4基因的转录调节作用机制。方法用RT-PCR及Real—timePCR检测HepG2．2．15细胞和HepG2细胞中DNAJB4在mRNA水平上的表达差异。构建DNAJB4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。结果在mRNA水平上，DNAJB4在HepG2．2．15细胞中的表达量是其在HepG2细胞中表达量的2．59倍，且二者有显著性差异（P〈0．01）。DNAJB4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是其对照组的2．28倍。转染HBs和HBc表达质粒组的相对荧光素酶活性分别是其对照组的2．11倍和1．77倍，而HBx、HBp表达质粒对荧光素酶活性没有影响。结论在HepG2．2．15细胞中，HBV能通过增强DNAJB4启动子活性增加DNAJB4转录水平的表达，其中HBs和HBc蛋白起主要作用。Objective This study was aimed to investigate the relationship between HBV and transcriptional expression level of DNAJB4. Methods RT-PCR and Real-time PCR were performed to compare DNAJB4 expression at mRNA level in HepG2 and HepG2. 2. 15 cell lines. The DNAJB4 promoter luciferase reporter plasmid, pGL3-Basie-DNAJB4-P, was constructed. HepG2 cells were transiently co-transfected with equal amount of pGL3-Basic-DNAJB4-P and HBV or HBx, HBs, HBc, HBp expression plasmid, respectively. Relative lueiferase activity was then detected. Results The amount of DNAJB4 expression at mRNA level in HepG2. 2. 15 cells was 2. 59 times higher than that in HepG2 cells. The relative lueiferase activity in HepG2 cells transiently co-transfected with pGL3-Basic-DNAJB4-P and HBV expression plasmid, was 2.28 times higher than that in control group. The relative lueiferase activity in HepG2 cells transfected with HBs or HBc expression plasmids was 2. 11 times or 1.77 times higher than that in control group, respectively. However, HBx and HBp expression plasmids had no effect on luciferase activity. Conclusion It was found that HBV up-regulates transcriptional level expression of DNAJB4 in HepG2.2. 15 cells via enhancing DNAJB4 promoter activity, where HBs, HBc proteins may play important roles.

关 键 词：乙型肝炎病毒 DNAJB4启动子 HEPG2.2.15细胞 基因表达与调控 

分 类 号：R373[医药卫生—病原生物学]