Study of RASSF1A expression and promoter demethylation in Hep-2 cell line.
- Author:
Jing YANG
1
;
Wenyue JI
;
Yarong QU
;
Lixia HE
;
Xudong ZHAO
Author Information
1. Department of Otolaryngology, Affiliated Shengjing Hospital of China Medical University, Shenyang, 110004, China.
- Publication Type:Journal Article
- MeSH:
Antimetabolites, Antineoplastic;
pharmacology;
Azacitidine;
analogs & derivatives;
pharmacology;
Cell Line, Tumor;
DNA Methylation;
Gene Expression;
Gene Silencing;
Genes, Tumor Suppressor;
Humans;
Hydroxamic Acids;
pharmacology;
Promoter Regions, Genetic;
Tumor Suppressor Proteins;
genetics
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2011;25(2):64-66
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of 5-Aza-dC and TSA to tumor suppressor gene RASSF1A expression and methylation level in Hep-2 cell line.
METHOD:Hep-2 cell line were cultured in vitro and handled with 5-Aza-dC and TSA. Detected RASSF1A expression and methylation level were detected before and after drug intervention using Realtime PCR and MSP.
RESULT:(1) Before intervention with drug, tumor suppressor gene RASSF1A was weakly expressed and methylated in Hep-2 cell line. (2) With the effect of 5-Aza-dC and TSA, the methylation of RASSF1A gene was reversed. And the effect of combination of 5-Aza-dC and TSA was similar with 5-Aza-dC alone. There was no obvious effect using TSA alone. (3) With the effect of 5-Aza-dC and TSA, the expression of RASSF1A was improved. And the effect of 5-Aza-dC was stronger than TSA. Synergetic effect was found when using 5-Aza-dC and TSA simultaneously.
CONCLUSION:In Hep-2 cell line, Promoter methylation of tumor suppressor RASSF1A may play a very important role in loss of gene expression, but it is not the only cause. 5-Aza-dC and TSA can improve RASSF1A expression by reversing DNA methylation and histone deacetylation.
