[Detection of p15 gene methylation or deletion status in different malignant cell lines by using hemi-nested methylation specific polymerase chain reaction].
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.
Lin
Fu-An
FA
Department of Hematology, The Affiliated Zhangzhou Municipal Hospital, Fujian Medical University, Zhangzhou 363000, China.
Ye
Bao-Guo
BG
Shen
Jian-Zhen
JZ
Zhou
Hua-Rong
HR
Fu
Hai-Ying
HY
Fan
Li-Ping
LP
D018899:CpG Islands / Q000235:genetics*; D050762:Cyclin-Dependent Kinase Inhibitor p15 / Q000235:genetics*; D019175:DNA Methylation; D017353:Gene Deletion; D015972:Gene Expression Regulation, Neoplastic; D019337:Hematologic Neoplasms / Q000235:genetics* / Q000473:pathology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D014407:Tumor Cells, Cultured
D050762:Cyclin-Dependent Kinase Inhibitor p15