肠道病毒71型3C蛋白表达后细胞蛋白质组差异分析

基础医学与临床 ›› 2011, Vol. 31 ›› Issue (6): 652-655.

肠道病毒71型3C蛋白表达后细胞蛋白质组差异分析

雷晓波,孙振民,刘新蕾,任丽丽,王健伟

中国医学科学院北京协和医学院病原生物学研究所

收稿日期:2011-03-09 修回日期:2011-04-11 出版日期:2011-06-05 发布日期:2011-06-06
通讯作者: 王健伟 E-mail:wangjw28@163.com
基金资助:
国家重大基础研究规划（973计划）项目资助

Proteomic Analysis of Differential Protein Expression in 293T Cells after Overexpression of Enterovirus 71 3C

Xiao-bo LEI¹,Zhen-min SUN²,Xin-lei LIU²,Li-li REN²,Jian-wei WANG³

1. Institute of Pathogen Biology, CAMS&PUMC
2.
3. Institute of Pathogen Biology, CAMS& PUMC

Received:2011-03-09 Revised:2011-04-11 Online:2011-06-05 Published:2011-06-06
Contact: Jian-wei WANG E-mail:wangjw28@163.com

摘要/Abstract

摘要： 目的利用蛋白质组学技术研究肠道病毒71型（EV71）编码3C蛋白对宿主细胞蛋白表达的影响。方法在293T细胞中瞬时过表达EV71 3C蛋白，应用双向荧光差异凝胶电泳（2D-DIGE）检测3C表达后293T细胞中蛋白的差异。选取表达水平有明显变化的蛋白用Western blot进行验证。结果在293T细胞中过表达3C蛋白后，可引起5个蛋白表达水平发生明显改变，其中4个蛋白表达上调，1个蛋白（HSPA4）表达下调。Western blot验证表明过表达3C可降低HSPA4的表达，HeLa细胞中HSPA4表达为对照的46.8%±10.1%（P＜0.05）。结论过表达EV71 3C蛋白可引起细胞内蛋白表达差异。

关键词: 肠道病毒EV71, 3C, 蛋白质组差异

Abstract: Objective To explore the effects of 3C protease on host cell proteins by proteomics techniques. Methods 293T cells were transfected with plasmid encoding EV71 3C, cell lysates were analyzed by using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mass spectrometry (MS) analysis. Down-regulated protein was confirmed by Western blot analysis. Results The expression of EV71 3C in 293T cells is confirmed. A total of 5 altered proteins were identified after 3C overexpression. Among them, expression of 4 proteins increased, while expression of 1 protein decreased. Down-regulated protein HSPA4 was confirmed by Western blot. HSPA4 expression was determined by performing densitometric analyses in HeLa cells. The results show that HSPA4 expression was 46.8%±10.1% (P＜0.05) compared to normal cells. Conclusions Overexpression of EV71 3C regulates the protein expression in 293T cells.

Key words: Enterovirus 71, 3C, proteomics differences

雷晓波孙振民刘新蕾任丽丽王健伟. 肠道病毒71型3C蛋白表达后细胞蛋白质组差异分析[J]. 基础医学与临床, 2011, 31(6): 652-655.

Xiao-bo LEI Zhen-min SUN Xin-lei LIU Li-li REN Jian-wei WANG. Proteomic Analysis of Differential Protein Expression in 293T Cells after Overexpression of Enterovirus 71 3C[J]. Basic & Clinical Medicine, 2011, 31(6): 652-655.