Effects Of MET Kinase Inhibitor BMS-777607 On Proliferation,apoptosis And Migration Of Tongue Squamous Cell Carcinoma And Its Related Mechanisms

Objective To investigate the effects of MET kinase inhibitor BMS-777607 on the proliferation,apoptosis and migration of tongue squamous cell carcinoma CAL27 cells,and to explore its related mechanisms.Methods Different concentrations of MET kinase inhibitor BMS-777607 were applied to tongue squamous cell carcinoma CAL27 cells,and the effect of BMS-777607 on the proliferation of tongue squamous cell carcinoma cells was detected by MTT method,clone formation experiment and Ed U cell imaging experiment.The effect of BMS-777607 on the apoptosis of tongue squamous cell carcinoma cells was observed by Heochst33342 staining.The effect of BMS-777607 on the mitochondrial membrane potential of tongue squamous cell carcinoma cells was detected by JC-1 staining.The effect of BMS-777607 on the migration of tongue squamous cell carcinoma cells was detected by scratch test and Transwell test.Western blot was used to detect the effects of BMS-777607 on the expression of proliferating protein AKT,p-AKT,and apoptosis-related protein Bcl-2,Cleaved caspase-3,Bax and PARP in tongue squamous cell carcinoma cells.Results MTT results showed that the cell proliferation rate of the experimental group decreased with the increase of drug concentration(P<0.01).The results of the clone formation experiment showed that the cells in the experimental group grew slowly compared with the control group,and the cells in the 10μ mol/L group were almost in the static phase,and no obvious clones were found.The cell proliferation rate in the experimental group decreased with the increase of drug concentration(P<0.01).The EDU cell imaging experiment showed that the cell nucleus number of the experimental group was significantly reduced under green light excitation,and the cell proliferation rate was decreased with the increase of drug concentration(P<0.01).The results of Hoechst33342 staining showed that the cells in the control group had regular morphology,intact nuclei and uniform staining.In the 5 μ mol/L group,most nuclei were stained with high luminance fluorescence,pyknosis and fragmentation in different degrees.The nuclei of most cells in the 10 μmol/L group showed high fluorescence staining,and pyknosis and fragmentation were more obvious.The results of JC-1 showed that the mitochondrial membrane potential of the control group showed strong red fluorescence,while the mitochondrial membrane potential of the experimental group showed green fluorescence,and the green fluorescence increased with the increase of drug concentration.The scratch test results showed that compared with the control group,the cell migration rate of the experimental group decreased with the increase of drug concentration,and there was a positive correlation between the cell migration rate and the action time(P<0.01).Transwell migration assay showed that the number of Transwell cells in the experimental group was lower than that in the control group,and the migration rate decreased with the increase of drug concentration(P<0.01).Western detection results showed that compared with the control group,the protein expression levels of Bcl-2 and p-AKT were significantly decreased in the experimental group,while the protein expression levels of Cleaved caspase-3,Bax and PARP were significantly increased in a concentration-dependent manner(P<0.01).There was no significant change in AKT expression level(P>0.05).Conclusion BMS-777607,as a MET kinase inhibitor,can inhibit the proliferation and migration of tongue squamous cell carcinoma CAL27 cells and promote their apoptosis,and the mechanism may be related to PI3 K /AKT pathway and mitochondrial pathway.