Study of RASSF1A expression and promoter demethylation in Hep-2 cell line / 临床耳鼻咽喉头颈外科杂志

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.