Effects of 5-Aza-Cde on DNA Methylation and Expression of hMLHl and MGMT Gene in Lung Cancer Cell Line A549/DDP
10.3969/j.issn.1671-7414.2015.03.024
- VernacularTitle:5-氮杂-2′脱氧胞苷对肺癌 A549／DDP 细胞hMLHl，MGMT 基因甲基化及其表达的影响
- Author:
Hong WANG
;
Lili LI
;
Jicai ZHANG
;
Bo GAO
;
Haijun LUO
- Publication Type:Journal Article
- Keywords:
methylation;
drug fast;
hMLH1;
MGMT;
5-Aza-Cde
- From:
Journal of Modern Laboratory Medicine
2015;(3):83-86
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of 5-Aza-2′-deoxycytidine (5-Aza-Cde)on DNA methylation and expression of hMLH1 and MGMT gene in the human lung cancer cell line A549/DDP.Methods A549/DDP cells were cultured with RPMI 1 640 medium and were treated with 5 μmol/L DNA methyhransferase inhibitor 5-Aza-Cde.Methylation-specific pol-ymerase chain reaetioll (MSP)was used to detect the promoter methylation state of the hMLH1 and MGMT gene.RT-PCR was used to detect the mRNA expression of hMLH1 and MGMT before and after treatment with 5-Aza-Cde,respectively. Results Before treatment with 5-Aza-Cde,hMLH1 and MGMT expressions were absent,and promoter hypermethylation of the hMLH1 and MGMT gene were detected in A549 cells.After treatment with 5-Aza-Cde,the promoter region of the hM-LH1 and MGMT gene exhibited a demethylation state,and their mRNA expressions were increased.Conclusion Promoter hypermethyhtion is amajor mechanism of hMLH1 and MGMT gene silencing in human lung cancer cells,and can be reversed by the demethylating agent 5-Aza-Cde,which can regulate the expressions of the hMLH1 and MGMT gene.
