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目的探讨丙型肝炎病毒（ＨＣＶ）核心蛋白对肝瘤细胞血管内皮生长因子（ｖａｓｃｕｌａｒ ｅｎｄｏｔｈｅｌｉａｌｇｒｏｗｔｈ ｆａｃｔｏｒ， ＶＥＧＦ）表达的影响。方法将 ＨＣＶ核心基因 ｃＤＮＡ插入真核表达载体 ＰＢＫ－ＣＭＶ的Ｈｉｎｄ Ⅲ和ＢａｍＨ １位点间，构建重组质粒ＰＢＫ－ＨＣＶｃ。再将重组质粒ＰＢＫ－ＨＣＶｃ和空载体分别导入肝癌细胞株ＨｅｐＧ２中，Ｇ４１８筛选，ＲＴ－ＰＣＲ、蛋白印迹鉴定ＨＣＶ核心蛋白表达。免疫组织化学，蛋白印迹检测ＶＥＧＦ蛋白表达；原位杂交， ＲＴ－ＰＣＲ检测ＶＥＧＦ ｍＲＮＡ。结果重组质粒ＰＢＫ－ＨＣＶｃ在ＨｅｐＧ２细胞中有稳定表达；表达ＨＣＶ核心蛋白的细胞ＨｅｐＧ２－Ｃ的ＶＥＧＦ在蛋白水平和ｍＲＮＡ水平较转染空载体的细胞ＨｅｐＧ２－ＣＭＶ明显升高。结论ＨＣＶ核心蛋白能激活ＶＥＧＦ表达，可能对肝细胞癌的生长具有促进作用。
Objective To investigate the effect of hepatitis C virus (HCV) core protein on the expression of vascular endothelial growth factor (VEGF) in hepatoma cells. Methods HCV core gene cDNA was inserted into the Hind Ⅲ and BamH 1 sites of the eukaryotic expression vector PBK-CMV to construct the recombinant plasmid PBK-HCVc. The recombinant plasmids PBK-HCVc and empty vector were respectively introduced into HepG2 hepatoma cell line and screened by G418. The expression of HCV core protein was identified by RT-PCR and Western blotting. Immunohistochemistry and Western blotting were used to detect the expression of VEGF protein. VEGF mRNA was detected by in situ hybridization and RT-PCR. Results The recombinant plasmid PBK-HCVc was stably expressed in HepG2 cells. The expression of VEGF in HepG2-C cells expressing HCV core protein was significantly higher than that in HepG2-CMV transfected with empty vector. Conclusion HCV core protein can activate VEGF expression and may promote the growth of hepatocellular carcinoma.