Asiatic Acid Inhibites The Proliferation,migration And Invasion Of Tca8113 Cells In Tongue Squamous Cell Carcinoma Through The P53/FAK Signaling Pathway

Objective To investigate the effect of Asiatic acid(AA)on proliferation,migration and invasion of human tongue squamous cell carcinoma line Tca8113 cells and to explore its anti-tumor molecular mechanism,so as to provide evidence for the clinical development of drugs for the treatment of tongue cancer.Methods1.The MTT colorimetric method was used to detect the effect of AA on the viability of Tca8113 cells,and a suitable drug concentration was selected for subsequent experiments.2.The effects of AA on Tca8113 cell migration and invasion were determined by cell scratch experiment and Transwell invasion experiment.3.Western blot was used to detect the effect of AA on the expression of tumor suppressor gene p53,focal adhesion kinase FAK and EMT-related factors E-cadherin,N-cadherin and Vimentin in Tca8113 cells.4.Immunofluorescence staining was used to detect the effect of AA on the expression of p53 and FAK proteins in Tca8113 cells.5.Tca8113 cells were treated with p53 inhibitor,FAK inhibitor and AA,and divided into control group,p53 inhibitor group,FAK inhibitor group,AA group,AA+p53 inhibitor group.The expression of p53 and FAK in Tca8113 cells was detected by Western blot.6.The expression of FAK in tissue samples of tongue squamous cell carcinoma patients was detected by immunohistochemistry and Western blot.Results1.AA significantly inhibited the proliferation of Tca8113 cells in a dose-dependent manner,and its IC50 value for 24 h was about 41.72μmol·L-1(P<0.05).2.Compared with the control group,AA treatment inhibited the migration and invasion of Tca8113 cells(P<0.01).3.Western blot results showed that compared with the control group,AA increased the expression of p53 and decreased FAK expression in Tca8113cells;Meanwhile,the expression of EMT-related proteins E-cadherin increased,while N-cadherin and Vimentin expression decreased,in a time-dependent manner(P <0.01).4.The fluorescence intensity was observed by confocal microscopy,and the results showed that AA treatment increased the expression of p53 and decreased the expression of FAK in Tca8113 cells.5.Western blot results showed that FAK inhibitors had no significant effect on the expression of p53.However,when p53 inhibitors were used,FAK increased significantly in both the p53 inhibitor group and the AA+p53 inhibitor group(P<0.01).6.Immunohistochemical results showed that FAK expression in poorly differentiated tongue squamous cell carcinoma tissues was higher,while that in well differentiated tongue squamous cell carcinoma was lower,and Western blot results showed that FAK expression in poorly differentiated tongue cancer tissues was significantly higher than that in well differentiated tongue cancer tissues(P<0.01).Conclusions AA can inhibit the proliferation,migration and invasion of Tca8113 cells in tongue squamous cell carcinoma.The mechanism may be related to up-regulation of p53 protein expression,down-regulation of FAK protein expression and inhibition of EMT.