Asiatic Acid Inhibited The Growth And Migration Of Tca8113 Cells In Tongue Squamous Cell Cancer By Inducing Apoptosis And Regulating EMT

Objective Through the effect of Asiatic acid（AA） on the proliferation,apoptosis and migration of human tongue squamous cell carcinoma Tca8113 and the effect of mouse transplanted tumor model,the anti-tongue cancer effect in vivo and in vitro and its anti-tumor mechanism were discussed.Methods 1.The proliferation inhibition effect of AA with different concentrations on Tca8113 cells was detected by MTT assay,and the appropriate concentration and time of action of the drug were screened according to the results for subsequent experiments.2.To observe the effect of AA on Tca8113 cell clonal formation by using clonal formation experiment.3.The influence of Hoechst 33342 staining on the morphology of nuclear apoptosis was observed,and the influence of flow cytometry（FCM） on the apoptosis rate was observed.4.The influence of AA on tca8113 cell cycle was analyzed by flow cytometry.5.The effect of AA on Tca8113 cell migration was detected by cell scratch assay.6.The effects of AA on the expression of apoptosis-related proteins bcl-2,Bax,cleaved caspase-3,p53,p21 and migration related proteins N-cadherin,E-cadherin and Vimentin in Tca8113 cells were detected by Western blot.7.By comparing the xenograft model of human tongue squamous cell carcinoma,the inhibitory effect of AA on tongue squamous cell carcinoma xenograft in mice was observed,and the apoptosis of tumor cells in tumor tissues was analyzed by TUNEL method.Results 1.AA can effectively inhibit the proliferation of Tca8113 cells in a dose-dependent manner,with IC50 value of 42.13μmol·L^-1 at 24h（P<0.05）.2.AA inhibited the formation of Tca8113 cell clones（P<0.05）.3.Compared with the control group,the number of apoptotic bodies,apoptotic cells and apoptosis rate increased in the AA treatment group（P<0.05）.4.Cell cycle was gradually blocked at G₂/M stage after AA treatment（P<0.05）.5.Compared with the control group,the migration and healing ability of Tca8113 cells after 24h of AA treatment was reduced（P<0.05）.6.Western blot results showed that the expression level of apoptosis-related protein bcl-2 in Tca8113 cells was reduced,and the expression levels of Bax,cleaved cappase-3,p53 and p21 were increased.At the same time,the expression of migration related proteins E-cadherin,a protein related to cell migration,was increased,while the expression of N-cadherin and Vimentin was decreased,showing a drug concentration dependence（P<0.05）.7.AA inhibited the growth of xenograft tumors in nude mice with tongue squamous cell carcinoma and promoted the apoptosis of cancer cells in tumor tissues（P<0.05）.Conclusions AA can inhibit Tca8113 tongue squamous carcinoma cell proliferation and migration of Tca8113 tongue squamous carcinoma cell apoptosis can be induced,and the above functions and AA concentration dependence and certain its promoting apoptosis mechanism may be associated with p53 pathway,its inhibition of migration may ectomesenchymal cells and Epithelial cells to change（Epithelial to Mesenchymal Transition,EMT） related factors.