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目的探讨乙肝病毒X蛋白(HBx)表达与肝细胞Period1基因低表达的关系及其机制。方法采用脂质体法将pcDNA3.1-HBx质粒转入CCL13细胞,采用甲基化聚合酶链式反应(PCR)、反转录PCR(RT-PCR)、蛋白质印迹法(western blot)检测CCL13细胞转染HBx质粒前后Period1基因启动子甲基化水平、mRNA及蛋白表达水平。结果 pcDNA3.1-HBx质粒转染CCL13细胞后在该细胞中稳定表达,转染HBx质粒后,Period 1在CCL13细胞启动子甲基化水平较转染空质粒CCL13细胞明显增高,其mRNA及蛋白质的表达水平均较后者明显下降。结论 HBx蛋白可通过上调肝细胞Period1启动子甲基化水平,促使其表达下调。
Objective To investigate the relationship between the expression of hepatitis B virus X protein (HBx) and the low expression of Period1 gene in hepatocytes and its mechanism. Methods The pcDNA3.1-HBx plasmid was transfected into CCL13 cells by lipofectamine. The expression of CCL13 was detected by reverse transcription polymerase chain reaction (RT-PCR) and western blot Period1 gene promoter methylation level, mRNA and protein expression level before and after HBx plasmid transfection. Results After pcDNA3.1-HBx plasmid was transfected into CCL13 cells, the expression of pcDNA3.1-HBx was stably expressed in the cells. After transfection with HBx plasmid, the promoter methylation level of Period 1 in CCL13 cells was significantly higher than that in CCL13 transfected cells. The mRNA and protein The expression level of the latter was significantly lower than the latter. Conclusion HBx can down-regulate the expression of Period1 promoter in hepatocytes.