结合消减杂交筛选肝癌细胞系中差异甲基化位点  

Scanning the Differential Methylation Sequences in Hepatocellular Carci-noma Cell Line by Subtractive Hybridization

作　　者：樊红[1] 程欲超[1] 赵主江[1] 权艳梅[1] 成建[1] 

机构地区：[1]东南大学"发育与疾病相关基因"教育部重点实验室

出　　处：《生物医学工程研究》2006年第3期154-157,共4页Journal Of Biomedical Engineering Research

基　　金：国家自然科学基金资助项目(30470950)

摘　　要：为探索有效分离两样品间差异甲基化片段的方法,以DNMT1(DNAmethyltransferase1,DNMT1)表达抑制前后的肝癌细胞系7721-sMT1和7721-pMT1为研究对象,结合消减杂交和磁珠分离的方法,在两细胞系全基因组DNA范围内进行扫描差异性甲基化片段;将这些片段进行生物信息学分析,发现所获38条差异甲基化片段分布于不同的染色体上,但集中于重复序列、基因的启动子区,此特点符合DNA甲基化位点分布规律。实验结果证明消减杂交结合磁珠分离的方法,能筛选出两样品间的差异甲基化片段,该方法的建立为筛选肿瘤细胞中特异性差异甲基化片段作为肿瘤诊断的标志物提供了可能性。To set up an efficient method of getting the differential methylation sequences between two samples,two cell lines 7721-pMT1 and 7721-sMT1 which was suppressed or not by DNMT1 siRNA in hepatocellular carcinoma cell line SMMC-7721 were investigated by substrative hybridization combined with magnetic beads.The differential methylation sequences were scanned in global genomic DNA and were analyzed by bioinformatics.38 differential methylation fragments sitting on different chromosome were belonged to repeat sequence family and promoter region of certain gene through the whole genomic DNA.These sequences were accord with the distribution ruler of methylation sequence in whole genomic DNA.The method can be used to catch the differential methylation sequences between two samples in whole genomic DNA.Maybe it is a potential tool to catch methylation sequences in tumor tissues as an epigenetics marker used to diagnosis.

关 键 词：消减杂交 DNA甲基化 差异甲基化片段 表观遗传学 CPG岛 DNMT 

分 类 号：R318[医药卫生—生物医学工程]