Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression

Chuanli LIU; Jinju MAO; Yuanyuan YE; Qin HU; Lijun ZHANG; Weixian CHEN

doi:10.16016/j.1000-5404.201904168

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Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression

Basic Medicine | 更新时间：2026-01-09

- Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression
- Journal of Third Military Medical University Vol. 41, Issue 18, Pages: 1744-1749(2019)
- 作者机构：
  
  400010 重庆，重庆医科大学附属第二医院检验科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(31600135);Key Project of Chongqing Health Commission(20141005)
- DOI：10.16016/j.1000-5404.201904168
  CLC： R373.21;R394.3;R341
- Received：11 April 2019，
  
  Revised：2019-05-23，
  
  Published：30 September 2019
- 稿件说明：
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Chuanli LIU, Jinju MAO, Yuanyuan YE, et al. Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression[J]. Journal of Third Military Medical University, 2019, 41(18): 1744-1749.
DOI：

Chuanli LIU, Jinju MAO, Yuanyuan YE, et al. Hepatitis C virus core protein interacts with zinc finger E-box binding homeobox 2 to inhibit E-cadherin expression[J]. Journal of Third Military Medical University, 2019, 41(18): 1744-1749. DOI： 10.16016/j.1000-5404.201904168.

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(

0.01)，而ZEB2的蛋白和mRNA表达水平明显增加(

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(

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(

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).

Methods

A 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.

Results

Compared with the control cells

the cells transiently and stably expressing HCV core protein both showed significantly decreased expression of E-cadherin (

0.01) and increased ZEB2 expression (

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 (

0.01)

while ZEB2 knockdown by RNA interference partly recovered the luciferase activity (

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.

Conclusion

HCV core protein increases the expression of ZEB2 and enhances its binding to E-cadherin promoter

and thus inhibits the expression of E-cadherin.

关键词

Keywords

references

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