半巢式甲基化特异性聚合酶链反应对肿瘤细胞株p15基因甲基化或缺失状态的检测.docVIP

半巢式甲基化特异性聚合酶链反应对肿瘤细胞株p15基因甲基化或缺失状态的检测 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：半巢式甲基化特异性聚合酶链反应对肿瘤细胞株p15基因甲基化或缺失状态的检测 1 文2：四种基因甲基化特异性PCR引物设计探讨 8 1 方 法 9 2 结 果 10 3 讨 论 11 参考文摘引言： 12 原创性声明（模板） 14 文章致谢（模板） 14 正文 半巢式甲基化特异性聚合酶链反应对肿瘤细胞株p15基因甲基化或缺失状态的检测 文1：半巢式甲基化特异性聚合酶链反应对肿瘤细胞株p15基因甲基化或缺失状态的检测 Detection of p15 Gene Methylation or Deletion Status in Different Malignant Cell Lines by Using Heminested Methylation Specific Polymerase Chain Reaction Abstract This study was aimed to investigate the methylation or deletion status of p15 gene in different malignant cell lines，and further to clarify their roles in the development and progression of malignant tumo. Heminested methylation specific polymerase chain reaction (hnMSP) was adopted to analyze p15 gene methylation or deletion status in 20 malignant tumor cell lines and mononuclear cells or normal cell lines in healthy people, as well as to evaluate its seitivity and specificity. The results showed that among all of the cell lines, Molt4,KG1,NCE,Raji,SMMC7221,CA46,SW480 and NCIH446 were partial methylated with CDKN2B gene，and its seitivity of detection of p15 gene methylation was up to ×10-5, also it had great specificity. Peripheral blood mononuclear cell(MNCs)from healthy volunteer,HL60,HepG2,293,HeLa,SGC7901,U266 and CEM were unmethylated; and K562,NB4,GMC,Jurkat seemed to have deletion or mutation of p15 gene. It is concluded that the incidence of p15 gene methylation or deletion in many tumou, especially malignant hematopathy, is frequent, they correlate with disease progression and prognosis. HnMSP is highly seitive and specific in analyzing p15 gene methylation, deserving in clinical application. Key words p15INK4B；gene methylation；gene deletion；heminested MSP J Exp Hematol 2007; 15(2):382-386 表观遗传学是指基于非基因序列改变所致基因表达水平变化，如DNA甲基化、组蛋白修饰和染色质重塑，在肿瘤发生发展中具有重要的作用。DNA甲基化被认为是肿瘤形成的重要机制之一，抑癌基因可因高度甲基化而失活，这种基因沉默调控与体细胞突变共同促进了肿瘤的发展。目前检测基因甲基化状态的方法有很多种［1-3］，概括起来可分为3组：①应用甲基化敏感的限制性内切酶方法。这种方法有可能因为酶切反应的不完