【摘要】 目的:微小RNA(microRNA)异常在肿瘤发生发展中起重要作用,本研究探讨microRNA mir219在肝细胞癌中的异常甲基化和异常表达,可能参与肝细胞癌的发生发展过程。方法:采用pyrosequence、TA克隆测序等方法检测正常肝组织、肝细胞癌细胞株和肝癌标本中mir219上游预测启动子区域甲基化,Taqman实时定量RT-PCR测定肿瘤组织表达。结果:与正常肝脏组织相比,肝癌细胞株、肝癌标本中mir219上游预测启动子区域中茎环区和上游1kb区域的CpG岛存在高度甲基化。肝癌细胞株、肝癌标本中mir219表达显著下调,与其异常甲基化相关。结论:肝癌中微小RNA mir219茎环序列上游1094bp区域存在启动子区异常甲基化,表达下调,提示mir219异常可能在肝细胞癌发生发展中起重要作用,是潜在的抑癌基因。更多还原

【Abstract】 Objective:To study the aberrant methylation in putative promoter region of MicroRNA mir219 and the methylation correlated expression down-regulation in HCC(hepatocellular carcinoma),and explore the potential role of mir219 in HCC.Methods:In HCC tumor sample and cell line,the methylation status of putative promoter region of mir219 by pyrosequence and TA sequencing were detected;and the expression detected by Taqman real time RT-PCR.Results:There were aberrant methylation in the CpG Island 1kb upstream and stem-loop sequence of mir219 in HCC,and the mir219 transcription was also significant down-regulated in HCC cell line.Conclusion:The 1094 bp upstream and stem-loop sequence was putative promoter region of mir219,which was aberrantly methylated in HCC and transcription down-regulated in HCC.It indicated its potential role in HCC development.更多还原