miR-125b对HBV复制及表达调控分析
The effect of miR-125b on HBV replication and expression in vitro
目的：研究miRNA-125b在乙肝病毒感染细胞模型中的表达变化及其与HBV相互作用。方法：miRNA芯片检测比较HBV稳定表达细胞系HepG2.2.15细胞与亲本细胞系HepG2细胞miRNA表达差异，采用实时定量RT-PCR检测miR-125b在HepG2.2.15及HepG2细胞中表达差异。HepG2.2.15细胞中过表达miR-125b，qPCR检测HBV DNA拷贝数，ELISA法检测HBsAg、HBeAg水平，Western blot检测HBc水平,southern blot 检测HBV复制中间体。干扰过表达miR-125b，检测HBV蛋白及DNA水平变化。结果：miRNA芯片检测共发:29条miRNA在HepG2.2.15细胞中表达上调，45条miRNA表达下调，进一步通过qRT-PCR检测显示，miR-125b在HepG2.2.15细胞中表达明显下调 (P<0.05)。在HepG2.2.15细胞过表达miR-125b，HBV DNA和蛋白表达水平均明显下调(P<0.05)；干扰过表达miR-125ba水平，HBV蛋白表达水平相应升高(P<0.05)。结论：miR-125b在HBV感染细胞模型中表达下调， miR-125b对HBV复制及蛋白表达具有抑制作用。
Objective：To investigate the effect of microRNA-125b (miR-125b) on the life cycle of hepatitis B virus (HBV) replication and expression. Methods: The miRNA expression profile differences were compared by miRNA array between HepG2 and HepG2.2.15 cells. MiR-125b was down-regulatedand in HepG2.2.15 cells. Overexpression miR-125b in HepG2.2.15, HBV protein and DNA level were tested by ELISA, quantification RT-PCR and western blot. Interfering ectopic miR-125b expression, HBV protein and DNA level were tested by ELISA, quantification RT-PCR and western blot. Results: Totally74 miRNAsexpression levels changed in HepG2.2.15 cells, including 29 up-regulated and 45 down-regulated. The expression of miR-125b in HepG2.2.15 cells was significantly down-regulated by RT-PCR (P<0.05).When miR-125b was over expressed in HepG2.2.15 cells, HBV replication and protein expression levels in HepG2.2.15 cells were down regulated comparing with control group (P <0.05); The inhibtory effect was antagonized when miR-125b antagonists was induced. Conclusions:This study confirmed that miR-125b could inhibit HBV replication and protein expression in vitro.
张祯祯、赵敏、刘泉波
基础医学分子生物学生物科学研究方法、生物科学研究技术
乙肝病毒、微小RNA、调控
Hepatitis B virus microRNA regulation
张祯祯,赵敏,刘泉波.miR-125b对HBV复制及表达调控分析[EB/OL].(2017-04-26)[2025-08-18].http://www.paper.edu.cn/releasepaper/content/201704-580.点此复制
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