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中华普通外科学文献(电子版)

中华普通外科学文献(电子版) ›› 2015, Vol. 09 ›› Issue (06) : 431 -436. doi: 10.3877/cma.j.issn.1674-0793.2015.06.005

所属专题： 文献；

论著

丙型肝炎病毒核心蛋白通过活化stat3通路促进肝癌细胞上皮间质转化的研究
周嘉嘉1, 陈汝福1,†(), 邓小耿1, 高文超1, 郑上游1, 程帝1, 周泉波1, 张杰1   
  1. 1. 510120　广州，中山大学孙逸仙纪念医院外科
  • 收稿日期:2015-09-07 出版日期:2015-12-01
  • 通信作者: 陈汝福
  • 基金资助:
    国家自然科学基金青年基金项目(81000889); 广东省自然科学基金重点资助项目(2014A030311047); 广东省科技计划项目(2014A020212094)

Hepatitis C virus core protein activating stat3 pathway and promoting epithelial-mesenchymal transition in hepatocellular cells

Jiajia Zhou1, Rufu Chen1,†(), Xiaogeng Deng1, Wenchao Gao1, Shangyou Zheng1, Di Cheng1, Quanbo Zhou1, Jie Zhang1   

  1. 1. Department of Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China
  • Received:2015-09-07 Published:2015-12-01
  • Corresponding author: Rufu Chen
  • About author:
    Corresponding author: Chen Rufu, Email:
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周嘉嘉, 陈汝福, 邓小耿, 高文超, 郑上游, 程帝, 周泉波, 张杰. 丙型肝炎病毒核心蛋白通过活化stat3通路促进肝癌细胞上皮间质转化的研究[J]. 中华普通外科学文献(电子版), 2015, 09(06): 431-436.

Jiajia Zhou, Rufu Chen, Xiaogeng Deng, Wenchao Gao, Shangyou Zheng, Di Cheng, Quanbo Zhou, Jie Zhang. Hepatitis C virus core protein activating stat3 pathway and promoting epithelial-mesenchymal transition in hepatocellular cells[J]. Chinese Archives of General Surgery(Electronic Edition), 2015, 09(06): 431-436.

目的

探讨丙型肝炎病毒核心蛋白（HCVc）对肝癌细胞信号转导子和转录激活子3（stat3）通路及上皮间质转化（EMT）的影响。

方法

将含HCVc基因序列的重组质粒pEGFP-N3-HCVc转染人肝癌细胞HepG2，Real-Time PCR和Western blotting检测HCVc、总stat3、磷酸化stat3（p-stat3）及EMT指标蛋白E-cadherin、Vimentin、Snail的表达，划痕实验及Transwell实验检测细胞迁移和侵袭情况。

结果

划痕实验及Transwell实验结果显示，过表达HCVc可增强HepG2细胞的迁移和侵袭能力；stat3通路抑制剂AG490处理可抑制HepG2细胞的侵袭能力。RT-PCR和Western blotting结果显示，过表达HCVc后，HepG2细胞中p-stat3、Vimentin及Snail在表达升高，E-cadherin表达下降；AG490处理可下调p-stat3、Vimentin及Snail表达，增强E-cadherin表达。

结论

HCVc可活化stat3通路促进肝癌细胞上皮间质转化，增强肝癌细胞的迁移和侵袭能力。

关键词: 病毒核心蛋白质类, 肝肿瘤, 肝炎, 丙型, stat3, 上皮间质转化
Objective

To investigate the role of hepatitis C virus core protein (HCVc) on the activation of stat3 pathway and regulation of epithelial-mesenchymal transition in hepatocellular cells, which might be involved in tumor metastasis of liver cancer.

Methods

Recombinant plasmid pEGFP-N3-HCVc containing HCVc gene was transfected to over-expressed HCVc in HepG2 cells. Real-Time PCR and Western blotting were used to detect the expression of HCVc, stat3, phosphorylated stat3 (p-stat3), E-cadherin, Vimentin and Snail. Wound healing test and Transwell assay was employed to detect cell migration and invasiveness.

Results

Wound healing test and Transwell assay showed that over-expression of HCVc in HepG2 cells promoted the cell migration and invasiveness. Meanwhile, AG490 treatment inhibited cell migration and invasiveness of HCVc over-expressing cells. RT-PCR and Western blotting analysis showed that expression of HCVc, p-stat3, Vimentin and Snail were increased and E-cadherin was decreased in HCVc over-expressing HepG2 cells. Treatment of AG490 on HCVc over-expressing cells could attenuate HCVc-induced up-regulation of p-stat3, Vimentin and Snail expression and down-regulation of E-cadherin.

Conclusion

HCVc activates stat3 pathway and promotes epithelial-mesenchymal transition in HepG2 cells, which may be associated with enhanced cell migration and invasion in liver cancer.

Key words: Viral core proteins, Liver neoplasms, Hepatitis C, stat3, Epithelial-mesenchymal transition
图1 Western blotting检测肝癌细胞HepG2-HCVc中HCVc蛋白的表达
图2 划痕试验检测转染HCVc后肝癌细胞HepG2-HCVc的迁移能力
图3 Transwell侵袭实验检测转染HCVc后肝癌细胞HepG2-HCVc的侵袭能力（200×)，左图为HepG2-Vector细胞穿过基底膜情况，中图为HepG-HCVc细胞穿过基底膜情况，右图为两组细胞侵袭细胞数对比
图4 Real-Time PCR（A）和Western Blotting（B）检测HepG2-HCVc细胞中E-cadherin、Vimentin及Snail mRNA和蛋白的表达
图5 Western Blotting检测HepG2-HCVc细胞中总stat3和p-stat3蛋白的表达
图6 Western Blotting检测AG490处理后HepG2-HCVc细胞中总stat3和p-stat3蛋白的表达
图7 Transwell侵袭实验检测AG490处理后HepG2-HCVc细胞的侵袭能力
图8 Real-Time PCR（A）和Western Blotting（B）检测AG490处理后HepG2-HCVc细胞中E-cadherin、Vimentin及Snail mRNA和蛋白的表达
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