Effects Of Deoxycytidine On DNA Methylation State And Expression Of MRNA And Protein Of P16 Gene In TSCC SCC-9 Cells

Objects:To Study the effects of deoxycytidine (5-aza-2 deoxycytidine, DAC) on DNA Methylation state and expression of mRNA and protein of p16 gene in human squamous lingual carcinoma SCC-9 cells in vitro, and to study on the anti-tumor effect and mechanism.Methods:The SCC-9 cells which were cultured in vitro were divided into four groups, group 0、1、2 and3. Group 1、2、3 were processed through the three graded concentrations of DAC (1×10-7mol/L,1×10-6mol/L.1×10-5mol/L) in DMEM medium, while DMEM culture medium of equal dosage without DAC as the group zero was given in the control group. Q-MSP was used to detect the state of methylation of the P16 in SCC-9 cells after cultured 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.Result:The hypermethylation and non-methylated in SCC-9 p16 gene 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 than that in 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.Conclusions:1.P16 gene of the human squamous lingual carcinoma SCC-9 cells may be reversed gene hypermethylation by DAC;2.The methylation status of P16 gene in SCC-9 cells may be reversed by the effect on demethylation of DAC, and promote the expression of mRNA and protein in P16 gene;3.DAC can be used as a gene drug in the treatment of tongue squamous cell carcinoma;...