巢式MS-PCR法检测恶性血液病细胞株APC基因启动子甲基化状态  

Detection of APC Gene Promoter Methylation in Hematological Malignant Cell Lines by Nested-Methylation Specific Polymerase Chain Reaction

作　　者：吴雪梅[1] 沈建箴[1] 喻爱芳[1] 范丽萍[1] 周华蓉[1] 付海英[1] 沈松菲[1] 吴淡森[1] 

机构地区：[1]福建医科大学附属协和医院血液病研究所,福建福州350001

出　　处：《中国实验血液学杂志》2009年第4期957-960,共4页Journal of Experimental Hematology

基　　金：福建省自然基金资助项目(编号C0540014);福建省百千万人才工程基金资助项目(编号303052801);福建医科大学教授基金(编号JS06080)资助项目

摘　　要：本研究旨在应用巢式甲基化特异性PCR(nMS-PCR)法检测10种恶性血液系统肿瘤细胞株APC基因启动子的甲基化状态,并探究其在肿瘤发生发展中的作用,同时筛选出APC基因启动子高甲基化的肿瘤细胞株,将其作为研究基因甲基化与表达关系的细胞模型。采用nMS-PCR扩增亚硫酸盐修饰后的10种肿瘤细胞株基因组DNA,检测其APC基因启动子区CpG岛的甲基化状态。结果表明,CA46、U266、Molt4、K562、HL-60、CEM、AKR、U937、Raji细胞的APC基因启动子区均未甲基化,而Jurkat细胞的APC基因启动子区出现甲基化。结论:用nMS-PCR可以准确地检测出恶性血液病细胞株APC基因的甲基化状态,该方法操作简便,可用于检测各种肿瘤细胞基因的甲基化状态。This study was aimed to investigate the effeciency of nested methylation specific polymerase chain reaction （nMS-PCR） for detecting the APC gene promoter methylation and to clarify the roles of methylation in genesis and development of hematologic malignancies, as well as to screen the hematologic malignant cell lines with hypermethylation of APC gene promoter to use as an ideal cell model for exploring the relationship between gen methylation and gene expression. The genome DNA of 10 cell lines modified with bisulfide was amplified and the methylation status of APC gene promoter was detected by using nMS-PCR. The results showed that the methylation of APC gene promoter was detected in Jurkat cells, while could not be detected in CA46, U266, Molt4, K562, HL-60, CEM, AKR, U937 and Raji cell lines. In conclusion, APC gene methylation in hematological malignant cell lines can be accurately detected by nMS-PCP method, which is simple method for detecting methylation status of various hematological malignant cell lines.

关 键 词：巢式甲基化特异性聚合酶链反应 恶性血液病细胞株 APC基因 基因甲基化 

分 类 号：Q503[生物学—生物化学] R733[医药卫生—肿瘤]