摘要
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p EGFP-N3-HCVc转染人肝癌细胞Hep G2,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Hep G2细胞的迁移和侵袭能力;stat3通路抑制剂AG490处理可抑制Hep G2细胞的侵袭能力。RT-PCR和Western blotting结果显示,过表达HCVc后,Hep G2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。
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 p EGFPN3-HCVc containing HCVc gene was transfected to over-expressed HCVc in Hep G2 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 overexpression of HCVc in Hep G2 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 Hep G2 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 epithelialmesenchymal transition in Hep G2 cells, which may be associated with enhanced cell migration and invasion in liver cancer.
出处
《中华普通外科学文献（电子版）》
2015年第6期431-436,共6页
Chinese Archives of General Surgery(Electronic Edition)
基金
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)
