Lin Fu-An, Ye Bao-Guo, Shen Jian-Zhen, Zhou Hua-Rong, Fu Hai-Ying, Fan Li-Ping
Department of Hematology, The Affiliated Zhangzhou Municipal Hospital, Fujian Medical University, Zhangzhou 363000, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2007 Apr;15(2):382-6.
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 tumors. 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 sensitivity 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 sensitivity of detection of p15 gene methylation was up to 1.0 x 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 tumours, especially malignant hematopathy, is frequent, they correlate with disease progression and prognosis. Hn-MSP is highly sensitive and specific in analyzing p15 gene methylation, deserving in clinical application.
本研究旨在探讨不同恶性细胞系中p15基因的甲基化或缺失状态，并进一步阐明其在恶性肿瘤发生发展中的作用。采用半巢式甲基化特异性聚合酶链反应（hn-MSP）分析20种恶性肿瘤细胞系以及健康人单核细胞或正常细胞系中p15基因的甲基化或缺失状态，并评估其敏感性和特异性。结果显示，在所有细胞系中，Molt-4、KG1、NCE、Raji、SMMC-7221、CA46、SW480和NCI-H446的CDKN2B基因呈部分甲基化，其检测p15基因甲基化的敏感性高达1.0×10（-5），且具有较高的特异性。健康志愿者外周血单个核细胞（MNCs）、HL-60、HepG2、293、HeLa、SGC7901、U266和CEM未甲基化；而K562、NB4、GMC、Jurkat似乎存在p15基因的缺失或突变。结论是，p15基因甲基化或缺失在许多肿瘤尤其是恶性血液病中的发生率较高，它们与疾病进展和预后相关。Hn-MSP在分析p15基因甲基化方面具有高度敏感性和特异性，值得临床应用。
