肺癌病人肿瘤组织DNA高甲基化片段的筛选  

Screening of Hypermethylated DNA Fragments in Tumor Tissue Derived from Patients with Lung Cancer

作　　者：刘晋祎 安倩[1] 张建军[1] 雷文东[1] 程书钧[1] 高燕宁[1] 

机构地区：[1]中国协和医科大学中国医学科学院肿瘤研究所肿瘤医院

出　　处：《Acta Genetica Sinica》2004年第4期389-394,共6页

基　　金：国家重点基础研究发展规划项目 (编号 :G19980 5 12 0 7);国家科技攻关计划项目 (编号 :2 0 0 2BA711A0 6)资助~~

摘　　要：关于DNA甲基化在肿瘤中的作用的研究 ,大多集中在研究已知的抑癌基因启动子区的异常高甲基化。而一些未知的参与肿瘤发生的基因也可能受甲基化调控 ,寻找这些与肿瘤相关的基因 ,对深入了解肿瘤发生的机制具有重要意义。利用甲基化敏感性随机引物PCR(Methylation SensitiveArbitrarilyPrimedPCR ,MS AP PCR) ,检查了肺癌组织中基因组范围内CpG岛高甲基化情况 ,分离到 8个高甲基化片段 (hypermethylatedDNAfragment,HMDF)。通过克隆、测序和Blast、NewCpGseek软件分析 ,发现所有的片段均为典型的CpG岛 ,有 4个片段与人 2、7、9、10号染色体上的同源性为 99%～ 10 0 % ,但只有 1个是已知的基因。进一步利用NeuralNetworkPromoterPrediction、TSSG和TSSW等软件对其余 7个片段可能的生物学意义进行了分析 ,结果有 4个片段是候选的启动子区 ,提示它们可能源于新基因。所获得的高甲基化片段可能是中国人肺癌发生过程中特有的表遗传学改变。The investigations on the role of DNA methylation in carcinogenesis have been mainly focused on promoter hypermethylation of tumor suppressor genes.As a number of genes associated with cancer development may be influenced by DNA methylation,identification of these genes is of great importance for understanding the epigenetic alteration in carcinogenesis.In this study,hypermethylated regions of genomic DNA from Chinese lung cancer patients were identified by a modified methylation-sensitive arbitrarily primed PCR (MS-AP-PCR).Eight hypermethylated DNA fragments (HMDF) were separated from a PCR product region between 300 and 500bp in size.After cloning,sequencing and searching with Blast and NewCpGseek programs,the result showed that all of them were typical CpG island sequences,four fragments had 99%～100% homology to regions on human chromosome 2,7,9 and 10,respectively,but only one revealed to be known gene.Neural Network Promoter Prediction,TSSG and TSSW programs were run to analyze possible functions of the rest 7 fragments,of which 4 were identified as candidate promoter regions,indicating that they might belong to new genes.The hypermethylated DNA fragments identified in this study might be specific epigenetic alterations in the Chinese lung cancer.

关 键 词：肺癌 DNA甲基化 表遗传学改变 甲基化敏感性随机引物PCR 

分 类 号：R734.2[医药卫生—肿瘤]