摘要
背景乙型肝炎病毒X蛋白(HBX)在乙型肝炎病毒复制及肝癌形成中扮演着重要的角色。研究HBX相关微小RNA(miRNAs)的差异表达有可能为调控乙型肝炎病毒复制、阐明肝癌发展进程及开展预后干预提供重要信息。目的探讨肝癌细胞中HBX相关miRNAs的差异表达及其在乙型肝炎病毒复制中的作用。方法选取2014年1—10月第二军医大学附属东方肝胆外科医院收治的13例肝癌患者的肝癌组织及距离肝癌组织>2 cm癌旁组织的新鲜石蜡包埋标本,采用微阵列技术分析HBX相关miRNAs的差异表达,实时荧光定量反转录聚合酶链式反应(RT-q PCR)法定量分析肝癌组织与癌旁组织中7个与肝脏功能相关的miRNAs及HBX mRNA表达水平,同时分析过表达hsa-miR-373或抑制hsa-miR-22对乙型肝炎病毒复制的影响。结果微阵列技术分析显示,HepG2-X细胞系和HepG2-CAT细胞系中有46种miRNAs差异表达,与HepG2-CAT细胞系相比,HepG2-X细胞系中20种miRNAs表达上调,26种miRNAs表达下调。肝癌组织与癌旁组织中hsa-miR-22、hsa-miR-373、hsa-miR-92、hsa-miR-155及HBX mRNA表达水平比较,差异均有统计学意义(P<0.05);肝癌组织与癌旁组织中hsa-miR-181a、hsa-miR-34a、hsa-let-7i表达水平比较,差异均无统计学意义(P>0.05)。Pearson相关性分析结果显示,hsa-miR-22表达水平与HBX mRNA表达水平呈正相关(r=0.415,P=0.018),hsa-miR-373、hsa-miR-92、hsa-miR-155表达水平与HBX mRNA表达水平呈负相关(r=-0.322,P=0.043;r=-0.369,P=0.028;r=-0.426,P=0.016);hsa-miR-181a、hsa-miR-34a、hsa-let-7i表达水平与HBX mRNA表达水平无直线相关关系(r=0.076,P=0.363;r=0.017,P=0.676;r=0.122,P=0.242)。第2天和第3天hsa-miR-22抑制剂组、hsamiR-373前体组HepG2.2.15细胞系及HepAD-38细胞系乙型肝炎病毒拷贝数较阴性对照组降低(P<0.05);第2天和第3天hsa-miR-373前体组Hep G2.2.15细胞系乙型肝炎病毒拷贝数较hsa-miR-22抑制剂组降低(P<0.05)。第2天hsa-miR-373前体组Hep AD
Background Hepatitis B virus X protein（ HBX）plays a very important role in HBV replication and hepatocarcinogenesis（ HCC） . Research on the differential expression of HBX - regulated miRNAs may provide important＆amp;nbsp;information for the regulation of HBV replication,the elaboration of the development process of liver cancer and intervention on prognosis. Objective To explore the differential expression of HBX - regulated miRNAs in HCC cells and its potential role in HBV replication. Methods We collected fresh specimens with paraffin embedding of HCC tissue and para - carcinoma tissue＆gt; 2 cm from HCC tissue from 13 HCC patients who were admitted into Eastern Hepatobiliary Surgery Hospital Affiliated to the Second Military Medical University from January to October in 2014. Microarry technology was used to analyze the differential expression of HBX - regulated miRNAs. Quantitative analysis was conducted to investigate the expression of seven miRNAs relevant with liver function and HBX mRNA in HCC tissue and para - carcinoma tissue by RT-qPCR method,and the influence of over - expression of hsa - miR - 373 and inhibiting - expression of hsa - miR - 22 on HBV replication was explored. Results The microarray results showed that there were a total of 46 differential expressions of miRNAs in HepG2 - X and HepG2 - CAT cell lines,and compared with HepG2 - CAT cell line,20 miRNAs expressions were higher and 26 miRNAs expressions were lower in HepG2 - X cell line. HCC tissue and para - carcinoma tissue were significantly different in the expression levels of hsa - miR -22,hsa - miR - 373,hsa - miR - 92 and hsa - miR - 155 and the expression level of HBX mRNA（P 〈 0. 05）;HCC tissues and para - carcinoma tissues were not significantly different in the expression levels of hsa - miR - 181a,hsa - miR - 34a and hsa- let - 7i（P ＆gt; 0. 05）. Pearson correlation analysis showed that the expression level of hsa - miR - 22 was postitvely correlated with the expression level of HBX mRNA（r = 0. 415,P = 0.
出处
《中国全科医学》
CAS
CSCD
北大核心
2016年第15期1786-1791,共6页
Chinese General Practice
基金
国家自然科学基金资助项目(91029723)
