Effects of deoxycytidine on DNA methylation state and expression of mRNA and protein of pl6 gene in human squamous lingual carcinoma SCC-9 cells / 实用医学杂志

ABSTRACT

Objective To study the effects of deoxycytidine (5-aza-2 deoxycytidine, DAC) on DNA Methylation state and expression of mRNA and protein of pl6 gene in human squamous lingual carcinoma SCC-9 cells in vitro. Methods The SCC-9 cells were divided into four groups, group 0, 1, 2 and 3 which processed using three gradients concentration of DAC. The group 0 without DAC was as the control group. Q-MSP was used to detect the state of methylation of the p16 in SCC-9 cells treated by DAC after 48 hours. Real-time fluorescence quantitative PCR was used to detect mRNA expression level changing of the p16 in SCC-9 cells treated by DAC. Immunohistochemical method was used to detect the expression of p16 protein. Results The hypermethylation and non-methylated p16 gene in SCC-9 was mixed with the results of Q-MSP. The results of Real-time PCR showed that mRNA expression of p16 in SCC-9 cells which treated by the different concentration of DAC for 48 hours was higher the control group. And difference was statistically significant (P < 0.05). The high expression of p16 protein was found in the experimental group with immunohistochemical method. Conclusion The p16 gene methylation states of SCC-9 may be suppresses and the recovery of mRNA and protein expression of p16 gene must be prompted by DAC.