肝细胞中HBV X基因的表达及其共培养的肝星状细胞的增殖和迁移

目的:研究乙型肝炎病毒X(hepatitis B virus X,HBV X)基因在乙型肝炎病毒致肝纤维化中的作用.方法:构建HBV X基因真核表达载体pHBV-X-IRES2-EGFP,将其转染人肝细胞HL-7702后分成2组,一组经G418筛选出稳定表达HBVX基因的肝细胞株(L02/x),另一组予瞬时转染48 h(L02/48x).Real-time PCR、Western blot鉴定2组细胞HBV X基因的表达.与转染空质粒和未转染的肝细胞组对照,将L02/x和L02/48x细胞分别与肝星状细胞(hepatic stellate cells,HSCs)共培养36 h,并检测各组HSCs增殖和迁移情况.结果:Real-time PCR和Western blot实验显示,转染pHBV-X-IRES2-EGFP载体的L02/x和L02/48x细胞均有HBV X基因的表达.与转染空质粒和未转染的肝细胞组对照,与L02/x和L02/48x细胞共培养的HSCs的增殖和迁移均显著增多.结论:HBV X基因在肝细胞中的表达可以促进HSCs发生增殖和迁移,从而在乙型肝炎病毒致肝纤维化过程中起重要作用.

Expression of the hepatitis B virus X gene in liver cells promotes the proliferation and migration of co-cultured hepatic stellate cells

AIM:To determine whether the hepatitis B virus X(HBV X) gene is involved in the pathogenesis of hepatitis B-related cirrhosis.METHODS:A eukaryotic expression vector containing the HBV X gene(pHBV-X-IRES2-EGFP) was constructed and transfected into HL-7702 cells.The transfected cells were divided into two groups.One group was selected with G418 and named L02/x,which could express the HBV X gene stably,and another group was transfected with pHBV-X-IRES2-EGFP for 48 h and named L02/48x.The expression of HBV X was detected by real-time PCR and Western blot.L02/x and L02/48x cells were then co-cultured with hepatic stellate cells(HSCs) for 36 h,and the proliferation and migration of HSCs were detected.RESULTS:Real-time PCR and Western blot analyses showed that L02/x and L02/48x cells could express HBV X.Compared to HSCs co-cultured with HL-7702 cells transfected with empty vector and non-transfected cells,the proliferation and migration of HSCs co-cultured with L02/x or L02/48x cells signif icantly increased.CONCLUSION:The expression of the HBV X gene in HL-7702 cells could promote the proliferation and migration of HSCs and may play an important role in the pathogenesis of hepatitis B virus-induced liver f ibrosis.