Detecting the methylation of p16INK4A in primary hepatocellular carcinoma using a nested bisulfite sequencing-methylation specific polymerase chain reaction
- VernacularTitle:甲基化特异性PCR在原发性肝细胞癌p16INK4A基因甲基化检测中的应用
- Author:
Bin YANG
;
Yingtang GAO
;
Zhi DU
;
Lei ZHAO
;
Wenqin SONG
- Publication Type:Journal Article
- Keywords:
Methylation;
Liver neoplasms
- From:
Chinese Journal of Laboratory Medicine
2001;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detection the methylation of p16INK4A in primary hepatocellular carcinoma, a nested bisulfite sequencing and methylation-specific polymerase chain reaction (BS-MSP) protocol was designed and used.Methods Bisulfite-modified DNA were amplified to evaluate the quality of templates with a pair of bisulfite sequencing primers in the first round of PCR, then subjected to methylation assay with corresponding methylation or unmethylation specific PCR primers.Representative PCR products were sequenced to confirm its correctness.Results 3 of 40 cases (7.5%) were failed to assay due to poor quality of templates, and 29 of 37 cases (78%) were detected p16INK4A methylation.Sequencing results confirmed that templates were correctly amplified.Conclusion BS-MSP technique might be valuable for methylation study on carcinogenesis and clinical assay.
