摘要目的 探讨HBV是否通过调控组蛋白甲基化酶SMYD3的表达参与肝癌细胞的恶性生物学行为.方法 通过向肝癌细胞HepG2转染HBx筛选出表达HBx蛋白的细胞株HepG2-HBx.用激光共聚焦定位细胞内HBx蛋白和SMYD3蛋白的表达;实时逆转录PCR、Western blot检测转染HBx前后HepG2细胞中SMYD3 mRNA和蛋白质的表达水平;流式细胞仪检测转染HBx前后HepG2细胞增殖、凋亡的变化情况.对数据进行单因素方差分析. 结果 HBx转染后HepG2细胞中SMYD3 mRNA和蛋白质表达水平明显上调(SMYD3 mRNA:0.18±0.05与0.98±0.15,F=37.240,P<0.05;SMYD3蛋白:0.28±0.03与0.58±0.06,F=21.042,P<0.05).转染HBx后HepG2细胞凋亡率下降(7.90%±0.42%与2.23%±0.14%,P<0.01),细胞增殖能力增强(28.46%±4.33%与36.46%±0.17%,P<0.01).结论 乙型肝炎病毒X蛋白能够上调HepG2细胞中SMYD3 mRNA和蛋白质表达水平;HBx可能通过SMYD3-组蛋白甲基化途径抑制HepG2细胞凋亡、促进细胞增殖.

abstractsObjective To explore the role SMYD3 and histone methylation in the carcinogenesis of HBV-related hepatocellular carcinoma (HCC). Methods HBx expressing plasmid was transfected into HepG2 cell, the localization of HBx and SMYD3 was detected by immunofluorescence, SMYD3 mRNA and protein were checked by real-time reverse transcription polymerase chain reaction and western blot, cell proliferation and apoptosis were detected by flow cytometry. Results After HBx transfection, HBx and SMYD3 protein were mainly localized in nucleus. HBx protein enhanced SMYD3 mRNA and SMYD3 expressions in HepG2. After HBx transfection, apoptosis of HepG2 was decreased, and cell proliferation was increased. Conclusions HBx may induce the expression of histone methyltransferase SMYD3, which in turn stimulates cell proliferation and blocks apoptosis.