摘要
目的探讨miR-583对乙型肝炎病毒(HBV)复制、表达的影响及相关作用机制。方法体外培养人肝癌细胞HepG2、人稳转HBV病毒的肝癌细胞HepG2.2.15,并将HepG2.2.15分为空白对照组、NC-siRNA组(转染NC-siRNA)及miR-583 siRNA组(转染miR-583 siRNA)。实时荧光定量PCR(qRT-PCR)法检测miR-583、DDX5 mRNA及HBV DNA表达,酶联免疫(ELISA)检测乙型肝炎病毒s抗原(HBsAg)、乙型肝炎病毒e抗原(HBeAg)表达水平,双荧光素酶法验证miR-583与DDX5的靶向关系。结果与HepG2细胞相比,HepG2.2.15细胞中miR-583表达水平显著升高(P<0.05),DDX5 mRNA表达水平显著降低(P<0.05)。与NC-siRNA组相比,miR-583 siRNA组HepG2.2.15细胞中miR-583表达水平显著降低(P<0.05),DDX5 mRNA表达水平显著升高(P<0.05),细胞上清液中HBV DNA、HBsAg、HBeAg表达水平显著降低(P<0.05)。Target ScanHuman网站预测及双荧光素酶实验结果均显示,miR-583与DDX5存在靶向调控关系。结论miR-583能促进HBV复制、表达,可能与靶向抑制DDX5表达有关,下调miR-583表达可抑制HBV复制、表达。
Objective To investigate the effects of miR-583 on the replication and expression of hepatitis B virus(HBV)and related mechanisms.Methods Human liver cancer cells HepG2 and human liver cancer cells HepG2.2.15,which are stably transfected with HBV virus,were cultured in vitro.HepG2.2.15 and divided into blank control group,NC-siRNA group(transfected with NC-siRNA)and miR-583 siRNA group(transfected with miR-583 siRNA).Real-time fluorescent quantitative PCR(qRT-PCR)method was used to detect the expression of miR-583,DDX5 mRNA and HBV DNA.The enzyme-linked immunoassay(ELISA)was used to detect the expression levels of hepatitis B virus antigen(HBsAg)and hepatitis B virus e antigen(HBeAg).The dual luciferase method was used to verify the targeting relationship between miR-583 and DDX5.Results Compared with HepG2 cells,the expression level of miR-583 in HepG2.2.15 cells was significantly increased(P<0.05),and the expression level of DDX5 mRNA was significantly reduced(P<0.05).Compared with the NC-siRNA group,the miR-583 expression level in HepG2.2.15 cells IN the miR-583 siRNA group was significantly reduced(P<0.05),the DDX5 mRNA expression level was significantly increased(P<0.05),and the expression levels of HBV DNA,HBsAg,and HBeAg in the cell supernatant were significantly reduced(P<0.05).Target ScanHuman website prediction and dual luciferase experiment results showed that miR-583 and DDX5 had a targeted regulatory relationship.Conclusion MiR-583 can promote HBV replication and expression,which may be related to the targeted inhibition of DDX5 expression.Down-regulating miR-583 expression can inhibit HBV replication and expression.
作者
黄学兰
王妍柏
王银锋
李想
HUANG Xuelan;WANG Yanbai;WANG Yinfeng;LI Xiang(Medical Experimental Center, General Hospital of Ningxia Medical University, Yinchuan 750004, China;Department of Neurology, General Hospital of Ningxia Medical University, Yinchuan 750004, China)
出处
《西部医学》
2022年第2期168-172,共5页
Medical Journal of West China
基金
宁夏自然科学基金项目(NZ16268)
宁夏医科大学校级重点项目(XZ2019007)。
