热休克蛋白90抑制剂17-AAG对乙型肝炎病毒复制的抑制作用

柯晓煜; 王秋景; 余源; 刘慎沛; 张春燕; 陈妍; 杨燕; 杨东亮

热休克蛋白90抑制剂17-AAG对乙型肝炎病毒复制的抑制作用

1. 华中科技大学同济医学院附属同济医院临床免疫研究室
2. 华中科技大学同济医学院附属同济医院实验医学研究中心

基金项目:

国家自然科学基金(30700701)；中央高校基本科研业务费专项资金(2011JC061)；

详细信息

中图分类号: R512.62
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出版历程
- 出版日期: 2012-06-20

17-AAG, an inhibitor of heat shock protein 90, down-regulates hepatitis B virus replication

Research funding:

摘要

摘要: 目的探讨热休克蛋白90（HSP90）是否参与肝细胞转化生长因子（TGF）β信号通路的活化,以及HSP90对乙型肝炎病毒（HBV）复制的影响。方法 HSP90抑制剂17-AAG作用HepG2细胞,提取细胞总RNA,实时定量RT-PCR检测TGFβ下游信号分子PAI-1的表达。HepG2细胞用17-AAG预处理2 h,同时用TGFβ或TβR抑制剂SB431542作用后,将HBV复制型质粒HBV1.3转染细胞,第4 d提取HBV核心颗粒,Southern Blot检测HBV复制中间体,ELISA检测上清HBsAg的表达。结果 17-AAG能够下调HepG2细胞PAI-1的表达,HepG2细胞内HBV复制中间体表达水平明显降低,HBsAg的表达亦受到抑制,但上调或阻断TGFβ信号通路对HBV复制影响不明显。结论 HSP90参与肝细胞内TGFβ信号通路的活化,其抑制剂17-AAG能够抑制HBV的复制与蛋白表达,但该抑制作用与TGFβ信号通路活化无关。
- HSP90热休克蛋白类 /
- 肝炎病毒,乙型 /
- 转化生长因子β
Abstract: Objective To investigate the potential involvement of the heat shock protein 90 (HSP90) in transforming growth factor-beta (TGFβ) signaling activation in hepatocytes, and to observe the effect of the HSP90 inhibitor, 17-AAG, on hepatitis B virus (HBV) replication.Methods HepG2 cells were treated with 17-AAG and total RNA was extracted to measure PAI-1 (SERPINE1) expression by real-time RT-PCR.Meanwhile, HepG2 cells were also transfected with the HBV1.3 HBV replicative plasmid after two hours of pretreatment with 17-AAG and TGFβ or SB431542.At day 4 post-transfection, HBV core particles were extracted and HBV replicative intermediates were detected by Southern blotting analysis.HBV surface antigen (HBsAg) was measured by ELISA.Results 17-AAG down-regulated mRNA expression of PAI-1 in HepG2 cells.HBV replication and HBsAg expression were inhibited upon 17-AAG treatment.However, activation of the TGFβ signaling pathway had no effect on HBV replication.Conclusion HSP90 is involved in the TGFβ signaling pathway during HBV infection.The HSP90 inhibitor, 17-AAG, can inhibit HBV replication and HBsAg expression through a TGFβ-independent signaling mechanism.
- HSP90 heat-shock-proteins /
- hepatitis B virus /
- transforming growth factor beta

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参考文献(12)

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