Effects Of Gossypol Acetic Acid On Protein Expressions Of ER,PR,C-fos,Dnmt1 Gene In Human Tongue Squamous Cell Carcinoma Cell Line Cal-27

Objective:1.To investigate the effects on proliferation and apoptosis in human tongue carcinoma cell line Cal-27 treated by GAA in vitro.2.To detect the effects on protein expressions of ER,PR,c-fos and Dnmtl gene in human tongue squamous cell carcinoma Cal-27 treated by GAA in vitro in order to provide theory and experimental base for prevention and therapy on carcinoma by changing the protein expressions of ER gene in human tongue squamous cell carcinoma Cal-27.Methods:1.The human tongue squamous cell carcinoma cell line Cal-27 were cultivated in vitro.The cells were treated with different concentrations of GAA.The morphological changes of Cal-27 cells treated with different concentrations of GAA were observed under an inverted microscope.2.The growth of human tongue squamous cell carcinoma cell line Cal-27 were detected at different concentrations of GAA for 48h and 72h through CCK8 assay,and the IC50 values of 48h and 72h GAA were calculated.3.To detect the apoptosis status changing in human tongue squamous cell carcinoma cell line Cal-27 before or after treated by different concentration of GAA for 48h and 72h through Flow Cytometry(FCM).4.To detect the protein expressions changing of ER,PR,c-fos and Dnmtl gene in human tongue squamous cell carcinoma cell line Cal-27 before or after being treated by different concentration of GAA for 48h and 72h through western blot assay.Results:1.The results of CCK-8 assay showed that the growth of human tongue squamous cell carcinoma cell line Cal-27 be inhibited after treated with 2.5?mol/L,5.0gmol/L and 10.0?mol/L GAA for 48h and 72h.In a certain range,the inhibition rate increases with the increase of concentration and duration of action.The IC50 values at 48h and 72h were 5.2?mol/L and 3.7?mol/L,respectively.2.The result of FCM showed that the apoptotic rates of human tongue squamous cell carcinoma cell line Cal-27 treated with 2.5?mol/L,5.0?mol/L and 00.?mol/L of GAA for 72h were higher than that of the control group(P<0.05).3.The results of Western-blot showed that the ER and c-fos protein expressions of human tongue squamous cell carcinoma cell line Cal-27 treated with 2.5?mol/L 5.0?mol/L and 10.O?mol/L of GAA for 48h and 72h were increased than the control group(P<0.05).Dnmtl and PR protein expressions were significantly lower than the control group(P<0.05).Conclusions:1.The GAA can inhibit the proliferation of human tongue squamous cell carcinoma Cal-27 cells in a concentration-dependent and time-dependent manner.2.The GAA can induce apoptosis in human tongue squamous cell carcinoma Cal-27 cells.In a certain range,the apoptosis rate increased with the increase of GAA concentration and time.3.GAA can decrease the expression of Dnmtl and PR protein in human tongue squamous cell carcinoma cell line Cal-27 and increase the expression of ER and c-fos protein.