Detection of promoter methylation of p16 gene in hematological malignant cell lines by nested methylation specific polymerase chain reaction.
- Author:
Hua-Rong ZHOU
1
;
Jian-Zhen SHEN
;
Hai-Yin FU
;
Bao-Guo YE
;
Li-Ping FAN
;
Fu-An LIN
Author Information
1. Fujian Institute of Hematology, Union Hospital, Fujian Medical University, Fuzhou 350001, China.
- Publication Type:Journal Article
- MeSH:
Base Sequence;
Cell Line, Tumor;
DNA Methylation;
Genes, p16;
HL-60 Cells;
Hematologic Neoplasms;
genetics;
pathology;
Humans;
K562 Cells;
Lymphoma;
genetics;
pathology;
Molecular Sequence Data;
Polymerase Chain Reaction;
methods;
Promoter Regions, Genetic;
genetics;
Sequence Analysis, DNA
- From:
Journal of Experimental Hematology
2006;14(2):375-378
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the efficiency of modified methylation-specific polymerase chain reaction i.e. nested methylation-specific polymerase chain reaction, used to detect the promoter methylation of p16 gene in six hematological malignant cell lines, and to explore the application in selection of hematological malignant cell lines with promoter hypermethylation, and make them to be an idel cell models for studying the relationship between gene methylation and expression. DNAs were denatured by NaOH and then were subjected to bisulfite modification and a nested-MSP was used to amplify the promoter region, nested MSP product of p16 gene promoter was analyzed and sequenced. The results showed that the hypermethylation of p16 gene was detected in CA46 and U266, however, Molt4, K562, HL-60 and Jurkat cell lines were unmethylated. In conclusion, p16 gene methylation in hematological malignant cell lines can be perfectly detected by nested-MSP method, which is simple, sensitive and specific for screening all kinds of hematological malignant cell lines with p16 gene methylated.
