Effects Of Gossypol Acetic Acid On Protein And The MRNA Expressions Of HMLH1and MGMT Gene In Human Tongue Carcinoma Cell Line Tca8113

Objective:To investigate the effects on protein expressions of hMLH1and MGMT gene in human tongue carcinoma cell line Tca8113treated by GAA in vitro;To detect the effects on mRNA expressions of hMLH1and MGMT gene in cell line Tca8113treated by GAA in vitro;To investigate the effects on mRNA expressions of DNMT1gene in human tongue carcinoma cell line Tca8113treated by GAA in vitro in order to discuss the mechanism of tumor suppression of GAA and provide the theory and experimental base for prevention and therapy on carcinoma tongue carcinoma cells treated by GAAMethods:To cultivate cell line Tca8113in vitro and detect the effects on protein expressions of hMLH1and MGMT gene treated by different concentration of GAA for24h,48h,72h through Western-blot; To study the interaction between hMLHl and MGMT gene in cell line Tca8113treated by GAA through co-immunoprecipitated; To investigate the effects on the mRNA expressions of hMLH1,MGMT and DNMT1gene in human tongue carcinoma cell line Tca8113treated by GAA for24h,48h through real-time fluorescence quantitative RT-PCR.Results:Compared with the control group, the result of Western-blot indicated that the protein expression of hMLHl and MGMT gene were increased treated by different concentration of GAA for24h,48h,72h (P<0.05);The result of co-immunoprecipitated showed that the protein expressions of hMLH1and MGMT gene in human tongue carcinoma cell line Tca8113has effect of interaction;The result of real-time fluorescence quantitative RT-PCR showed that the mRNA expression of hMLH1and MGMT gene in cell line Tca8113were increased after GAA treatment in a dose-dependent way(P<0.05). Meanwhile, the mRNA expression of DNMT1was reduced (P<0.05)Conclusion:The expression of protein and mRNA were reduction or missing in hMLH1and MGMT gene in human tongue carcinoma cell line Tca8113; The expression of protein and mRNA in hMLH1and MGMT gene in cell line Tca8113were increased after different concentration of GAA treatment. It indicated that GAA could reactivate these genes and upgrade its expression, which play its role as a suppressor on the potential tumor treatment target; The mRNA expression of DNMT1in hMLH1and MGMT gene in Tca8113cells was reduced after GAA treatment in a dose-dependent way. It indicated that GAA could downgrade the activation of DNMT1, which maybe one of the mechanisms of tumor suppression effect of GAA.