Replication of hepatitis B virus induces down-regulation of Arid2 expression in hepatoma cells
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Replication of hepatitis B virus induces down-regulation of Arid2 expression in hepatoma cells
Journal of Chongqing Medical UniversityVol. 40, Issue 3, Pages: 344-349(2015)
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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
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Published:2015,
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Li Zhi, Chen Zhen, Duan Yujie, et al. Replication of hepatitis B virus induces down-regulation of Arid2 expression in hepatoma cells. [J]. Journal of Chongqing Medical University 40(3):344-349(2015)
DOI:
Li Zhi, Chen Zhen, Duan Yujie, et al. Replication of hepatitis B virus induces down-regulation of Arid2 expression in hepatoma cells. [J]. Journal of Chongqing Medical University 40(3):344-349(2015)DOI:
Replication of hepatitis B virus induces down-regulation of Arid2 expression in hepatoma cells
目的:初步研究乙型肝炎病毒(hepatitis B virus,HBV)感染及其复制对肝癌细胞中抑癌基因Arid2(AT-rich interactive domain 2)表达的影响。方法:采用携带HBV基因组1.1倍体的复制型重组腺病毒Ad-HBV1.1感染Huh7细胞,或者四环素诱导的HBV复制细胞模型HepAD38细胞,观察HBV瞬时或稳定复制细胞模型中Arid2的表达变化;进一步采用HBV各元件乙型肝炎病毒S蛋白(hepatitis B virus S protein,HBs)、乙型肝炎病毒DNA聚合酶(HBV-pol)、乙型肝炎病毒核心蛋白(hepatitis B virus core protein,HBc)、乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)的过表达质粒转染Huh7细胞,明确病毒自身编码蛋白对Arid2表达的调控作用。在上述细胞模型中,运用Southern-blot、ELISA、real-time PCR的方法验证HBV的感染与复制情况,通过RT-PCR、Western blot检测肝癌细胞抑癌基因Arid2表达水平的变化。结果:HBV瞬时转染或稳定复制细胞模型中,Ad-HBV1.1感染的Huh7细胞mRNA和蛋白表达水平明显降低,Western blot结果显示Arid2蛋白表达水平降低了46.10%(P<0.05);在HBV稳定复制的HepAD38 细胞中,Arid2蛋白表达水平降低了37.63%(P<0.05)。转染HBV 4种病毒编码蛋白过表达质粒的Huh7细胞中,过表达 HBs、HBx组较其他实验组Arid2表达水平明显降低,Western blot结果显示Arid2 蛋白表达水平在HBs组降低了54.55%(P<0.05),在HBx组降低了46.38%(P<0.05)。结论:HBV的感染和复制导致肝癌细胞中Arid2的表达水平下调,Arid2基因的表达与HBV复制水平呈负相关,其中HBs、HBx对Arid2的下调作用较明显。
Abstract
Objective:To investigate the effects of hepatitis B virus replication on the expression of Arid2 in hepatoma cells. Methods:The HBV-stable or transient expression cell models were used to evaluate the regulatory effects of HBV replication on Arid2 expres-sion. For stable replication model,HepAD38 cells were used to express HBV under the control of inducible tetracycline promoter. For transient expression system,Huh7 cells were infected with the replicative recombinant adenovirus Ad-HBV1.1 carrying 1.1 copies of HBV genome. Southern-blot,ELISA and real-time quantitative PCR were employed to confirm HBV replication efficiency in hepatoma cells. The expression levels of Arid2 gene were determined by RT-PCR and Western blot. To further identify which of the viral-en-coded proteins implicated in the regulation of Arid2 expression,Huh7 cells were transfected with hepatitis B virus S protein(HBs),HBp,hepatitis B virus core protein(HBc) or hepatitis B virus X protein(HBx)-expression plasmids,and Arid2 expression level were determined. Results:HBV viroloigal marker and replicative intermediates were detected both in HBV-stably or transient expression system,indicating that HBV-stably or transiently expression cell model can robustly support HBV replication. The expression levels of Arid2 gene were significantly down-regulated both in the HBV stable replication and transient replication cell model. The protein expression level of Arid2 was decreased by 46.10% in Huh7 cells transfected with Ad-HBV 1.1 and by 37.63% in HepAD38 cells(P<0.05). Among the four viral-encoded expression plasmids,HBV surface antigen and HBx significantly repressed Arid2 mRNA and protein expression,with inhibitory effects of 54.55% and 46.38%(P<0.05) in protein level by HBs and HBx respec-tively. Conclusion:HBV replication represses Arid2 expression in hepatoma cells. Among viral-encoded proteins,HBs and HBx may play important roles in HBV-induced Arid2 repression.
关键词
乙型肝炎病毒Arid2乙型肝炎病毒S蛋白乙型肝炎病毒X蛋白
Keywords
hepatitis B virusArid2hepatitis B virus S proteinhepatitis B virus X protein
Effect of hepatitis B virus replication on the expression of inhibitor of differentiation family in hepatocytes
A preliminary investigation on the incidence of HBV-associated B-NHL and its clinical characteristics
Quantification of intrahepatic cccDNA,DNA,serum HBV DNA and its relationship with HBeAg in hepatitis B virus-related hepatocellular carcinoma
S gene variations of hepatitis B virus in children with vaccination failure
Association between intrahepatic levels and mutation of hepatitis B virus covalently closed circular DNA in hepatitis B virus-related hepatocellular carcinoma
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