丙型肝炎病毒核心蛋白对肝癌细胞stat3通路及细胞周期蛋白cyclin D1的调控作用  

Hepatitis C Virus Core Protein Activates stat3 Pathway and Regulates Cell Cycle Protein Cyclin D1 Expression in Hepatocellular Cells

作　　者：唐晶[1] 周嘉嘉[1] 邓小耿[1] 程帝[2] 张杰[1] 伍耀豪[1] 曾乐祥[1] 邱荣林[1] 陈汝福[1] 

机构地区：[1]中山大学孙逸仙纪念医院外科,广东广州510120 [2]中山大学附属第一医院黄埔院区肿瘤科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2013年第4期498-504,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30872485;81000889)

摘　　要：【目的】探讨丙型肝炎病毒核心蛋白(HCVc)是否调控肝癌细胞中stat3通路及细胞周期蛋白cyclin D 1的表达,参与肝癌细胞的恶性生物学行为。【方法】将含HCVc基因的真核表达质粒pEGFP-N3-HCVc瞬时转染人肝癌细胞HepG2使HCVc过表达,用siRNA-HCVc敲除HepG2细胞中HCVc的表达,荧光显微镜检测绿色荧光蛋白(GFP)的表达,Western blot检测HCVc、总stat3(stat3)、磷酸化stat3(p-stat3)及cyclin D1蛋白的表达;流式细胞术及MTT法检测细胞周期变化及细胞增殖情况。【结果】Western blot结果显示,通过瞬时转染而过表达HCVc的HepG2细胞中,HCVc、p-stat3及cyclin D1的蛋白水平高于空白质粒转染组和未转染组;siRNA-HCVc敲除HCVc表达后,HCVc、p-stat3和cyclin D1蛋白的表达下调;酪氨酸磷酸化抑制剂AG490(JAK2特异性拮抗剂)处理可下调过表达HCVc的HepG2细胞中p-stat3和cyclin D1蛋白的表达。流式细胞术显示过表达HCVc可促进细胞向G 2/M期进展。MTT法结果显示过表达HCVc使HepG2细胞增殖能力增加。【结论】HCVc可活化HepG2细胞中stat3通路,调控细胞周期蛋白cyclin D1的表达,促进细胞周期进展及细胞增殖,可能与肝癌的发展有关。[ Objective ] To investigate the role of hepatitis C virus core protein （HCVc） on the activation of stat3 pathway and regulation of cell cycle protein cyclin D1 expression in hepatocellular cells, which might involve in the malignant behavior of liver cancer cells. [Methods] Recombinant plasmid pEGFP-N3-HCVc containing HCVc gene was transiently transfected to overexpress HCVc in HepG2 cells, and siRNAs targeting HCVc was used to knockdown the expression of HCVc in HepG2 cells. Fluorescence microscopy was used to detect green fluorescent protein （GFP） expression, Western blot analysis aws used to detect HCVc, stat3, phosphorylated stat3 （p-stat3） and cyclin D1 protein expression. Flow cytometry and MTY assay was employed to detect cell cycle and cell proliferation. [Results] Western blot analysis showed that HCVc, p-STAT3, and cyclin D1 protein level were higher in HCVc transiently transfected HepG2 ceils than the blank plasmid transfected group and untransfected group, knockdown of HCVc expression by siRNAs can down-regulate the expression of p-stat3 and cyclin D1. Treatment of AG490 can downregulate the expression of p-stat3 and cyclin D1 in HCVc transfected HepG2 cells. Flow cytometry showed that overexpression of HCVc promote the progression of cell cycle into G2/M phase. MTr assay showed that overexpression of HCVc increases the proliferation of HepG2 cells. [Conclusions] HCVc activate stat3 pathway and regulates cell cycle protein cyclin D1 expression to promote cell cycle progression and cell proliferation in HepG2 cells, which may be associated with the development of liver cancer.

关 键 词：丙型肝炎病毒核心蛋白 肝癌 STAT3 CYCLIN D1 

分 类 号：R657.3[医药卫生—外科学]