Influnece of hepatitis B virus X protein on Period 1 expression and methylation level of CCL13 cells
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摘要: 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。Abstract: Objective To explore the relationship between hepatitis B virus (HBV)X pro-tein and inhibited expression of CCL13 cells Period 1.Methods The pcDNA3.1-HBx plasmid was transfected into CCL13 cells by Lipofectamine in vitro.Period1 expression and its promoter methylation level in CCL13 and CCL13-HBx were detected by treatments of reverse transcription PCR,western blot and methylation-specific PCR.Results The human liver cell lines CCL13-HBx contained stable HBx gene expression.Compared to CCL13,methylation level of period1 promoter increased in CCL13-HBx,and expression level of its mRNA and protein decreased.Conclusion Hepatitis B virus X pro-tein can decrease period1 expression in CCL13 by up-regulating its promoter methylation level.
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Keywords:
- liver /
- hepatitis B virus X protein /
- gene promoter /
- methylation
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