[1]刘传丽,毛金菊,叶媛媛,等.HCV核心蛋白与ZEB2的相互作用抑制E-cadherin的表达[J].第三军医大学学报,2019,41(18):1744-1749.
 LIU Chuanli,MAO Jinju,YE Yuanyuan,et al.Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression[J].J Third Mil Med Univ,2019,41(18):1744-1749.
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HCV核心蛋白与ZEB2的相互作用抑制E-cadherin的表达(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第18期
页码:
1744-1749
栏目:
基础医学
出版日期:
2019-09-30

文章信息/Info

Title:
Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression
作者:
刘传丽毛金菊叶媛媛胡琴张利军陈维贤
重庆医科大学附属第二医院检验科
Author(s):

Department of Clinical Laboratory, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

关键词:
丙型肝炎病毒核心蛋白肝细胞癌
Keywords:
hepatitis C virus core protein hepatocellular carcinoma
分类号:
R373.21; R394.3; R341
文献标志码:
A
摘要:

目的 探讨丙型肝炎病毒(hepatitis C virus,HCV)核心蛋白通过与E盒结合锌指蛋白2(zinc finger E-box binding homeobox 2,ZEB2)相互作用抑制E-cadherin表达的分子机制。方法 转染HCV核心蛋白真核表达质粒至HepG2细胞,并通过G418筛选构建稳定表达HCV核心蛋白的细胞株,以未转染HCV核心蛋白真核表达质粒的细胞作为对照组,采用RT-PCR和Western blot观测核心蛋白对ZEB2和E-cadherin表达水平的影响;采用荧光素酶报告系统分析核心蛋白对E-cadherin启动子活性的调控作用;通过免疫共沉淀及染色质免疫共沉淀技术检测核心蛋白与ZEB2的相互作用及与E-cadherin 启动子的结合情况。结果与对照组比较,在表达HCV核心蛋白的HepG2细胞中,E-cadherin的蛋白和mRNA表达水平显著降低(P<0.01),而ZEB2的蛋白和mRNA表达水平明显增加(P<0.05)。转染HCV核心蛋白的HepG2细胞中,E-cadherin启动子介导的荧光素酶活性显著降低(P<0.01),当利用RNA干扰技术敲减ZEB2后,E-cadherin启动子介导的荧光素酶活性明显恢复(P<0.05)。免疫共沉淀实验显示,HCV核心蛋白可与ZEB2结合,染色质免疫共沉淀实验表明,HCV核心蛋白增强了ZEB2与E-cadherin启动子的结合。结论 HCV核心蛋白可增强ZEB2的表达并促进其结合至E-cadherin启动子,从而抑制E-cadherin的表达。

Abstract:

Objective To investigate the molecular mechanism by which hepatitis C virus (HCV) core protein represses the expression of E-cadherin by interacting with zinc finger E-box binding homeobox 2 (ZEB2). MethodsA HepG2 cell line stably expressing HCV core protein was constructed by transfecting the cells with pCore-3FLAG and G418 screening. The expression levels of ZEB2 and E-cadherin in the transfected cells were examined with RT-PCR and Western blotting, and the promoter activity of E-cadherin was assayed using a dual luciferase system. The interaction of HCV core protein with ZEB2 was detected by immunoprecipitation, and the binding of ZEB2 with E-cadherin promoter was determined by chromatin immuoprecipitation. ResultsCompared with the control cells, the cells transiently and stably expressing HCV core protein both showed significantly decreased expression of E-cadherin (P<0.01) and increased ZEB2 expression (P<0.05) at both the protein and mRNA levels. HepG2 cells transfected with HCV core protein exhibited obviously lowered luciferase activity driven by E-cadherin promoter (P<0.01), while ZEB2 knockdown by RNA interference partly recovered the luciferase activity (P<0.05). Immunoprecipitation results demonstrated that HCV core protein could bind directly to ZEB2. Chromatin immunoprecipitation assay showed that the presence of HCV core protein obviously enhanced the binding of ZEB2 to E-cadherin promoter. ConclusionHCV core protein increases the expression of ZEB2 and enhances its binding to E-cadherin promoter, and thus inhibits the expression of E-cadherin.

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更新日期/Last Update: 2019-09-21