miRNA130a调控HCV复制的机制研究  

The Study on the Mechanism of MiRNA130a Regulating HCV Replication

作　　者：高晓洁 王倩 GAO Xiao-jie;WANG Qian(Department of Laboratory Medicine,Central Hospital of Hami,Hami 839000,China)

机构地区：[1]新疆哈密市中心医院检验科

出　　处：《标记免疫分析与临床》2019年第9期1556-1559,共4页Labeled Immunoassays and Clinical Medicine

摘　　要：目的探讨miRNA130a调控HCV复制的作用与机制。方法将Huh7.5.1细胞随机分为4组,正常组不予任何处理,感染组、实验组、NC组建立HCV感染模型;实验组在进行HCV感染前2 h在Huh7.5.1细胞内转染MiRNA130a mimic 100 nmol/L;NC组在HCV感染前2 h,加入等体积的无效miRNA。qRT-PCR法检测miRNA表达与HCV复制情况,CCK-8法检测细胞存活率,双染法检测细胞凋亡,Western blotting法检测细胞蛋白表达。结果感染组的MiRNA130a表达量低于正常组,实验组高于感染组,差异有统计学意义(P<0.05)。感染组的细胞存活率低于正常组,实验组的细胞存活率高于感染组;感染组的HCV复制水平高于正常组,实验组的HCV复制水平低于感染组,差异都有统计学意义(P<0.05)。感染组、实验组、NC组的细胞凋亡率高于正常组(P<0.05),感染组的细胞凋亡率最高(P<0.05)。感染组的PTEN、AKT蛋白表达水平高于正常组,实验组的PTEN、AKT蛋白表达水平低于感染组,差异有统计学意义(P<0.05)。结论MiRNA130a可抑制PTEN/PI3K/AKT信号通路,抑制HCV RNA的复制,降低细胞凋亡水平,提高细胞存活率。Objective To investigate the role and mechanism of miRNA130a in regulating HCV replication.Methods Huh7.5.1 cells were randomly divided into 4 groups:the normal group received no treatment,the infection group were given the miRNA130a treatment.The infected group,experimental group and NC group were used to establish the HCV infection model.The experimental group were transferred with MiRNA130a mimic 100 nmol/L 2 h before HCV infection;the NC group were given the equal volume of null miRNA.The miRNA expression and HCV replication were detected by qRT-PCR,the cell viability was detected by CCK-8 method,apoptosis were detected by double staining,and cell protein expression were detected by western blotting.Results The expression of miRNA130a in the infected group was lower than that in the normal group,and the experimental group higher than the infected group(P<0.05).The cell survival rate of the infected group was lower than that of the normal group,and the experimental group higher than that of the infected group.The HCV replication level of the infected group was higher than that of the normal group,and the HCV replication level of the experimental group was lower than that of the infected group(P<0.05).The apoptosis rates of the infected group,experimental group and the NC group were higher than that of the normal group(P<0.05),in which the infected group was the highest(P<0.05).The expression levels of PTEN and AKT proteins in the infected group were higher than those in the normal group.The expression levels of PTEN and AKT proteins in the experimental group were lower than those in the infected group(P<0.05).Conclusion miRNA130a can inhibit PTEN/PI3K/AKT signaling pathway,inhibit HCV RNA replication,reduce apoptosis and increase cell viability.

关 键 词：miRNA130a PTEN/PI3K/AKT信号通路 HCV复制 细胞凋亡 

分 类 号：R73[医药卫生—肿瘤]