乙肝患者HBV感染相关特异性miRNA筛选及其功能研究

目的寻找乙型肝炎病毒特异性的miRNA,并在体内验证找到的miRNA的功能,从而为阻断乙肝进一步发展提供新方法。方法利用miRNA芯片分析3对正常肝脏组织与慢性乙型肝炎组织的miRNA,寻找两者之间差异表达的miRNA。用实时定量聚合酶链反应(Real-time PCR,RT-PCR)在更多的组织标本中验证差异表达,确定有意义的miRNA。在鼠病毒性肝炎模型中通过静脉注射有意义的miRNA或anti-miRNA进行干预,同时刺激向肝癌方向转变,观察其病毒复制情况。结果与正常肝脏组织相比,慢性HBV病毒感染组织中表达上调的miRNA有8个,表达下调的miRNA有16个。miR-143在肝癌组织中的表达降低(P=0.0029);miR-520d-5p在肝癌组织中的表达提高(P=0.0320)。结论 miRNA在乙型肝炎的发生、发展中起重要作用,其功能的失调可能导致其最后发展成为肝硬化甚至肝癌,并且可以影响乙型肝炎治疗效果。

Screening and functional study of specific miRNA of HBV infection in hepatitis B patients

Objective To find the specific miRNA of hepatitis B virus and verify the functions of the miRNA in vivo,so as to provide a new method for blocking the further development of hepatitis B. Methods We used miRNA chips to analyze the miRNA of 3 pairs of normal liver tissues and chronic hepatitis B tissues, and found the different expression of miRNA. Real-time quantitative polymerase chain reaction (RT-PCR) was used to verify the different expression in more tissue samples to identify significative miRNA. Intervention by intravenous injection of the significative miRNA or anti-miRNA in the viral hepatitismodels of rat was done. At the same time, we stimulatedthem to turn into liver cancerand then observed the replication of the virus. Results Compared with normal liver tissue, 8 miRNAs were up-regulated in chronic HBV-infected tissues, and 16 miRNAs were down-regulated. The expression of miR-143 was decreased in hepatocellular carcinoma(P=0.0029), and the expression of miR-520d-5p in hepatocellular carcinoma was increased (P=0.032). Conclusion miRNA plays an important role in the incidence and development of hepatitis B. The dysfunction of it may lead to the final development of liver cirrhosis and even liver cancer, and it can affect the therapeutic effect of hepatitis B.