Effects Of Aurora Kinase Inhibitor VX-680 On Proliferation,Adhesion,Migration And Anoikis Of Human Esophageal Carcinoma Cells

Objective: Investigate the effect of Aurora kinase inhibitor VX-680 on proliferation,apoptosis,adhesion,migration and anoikis of human esophageal carcinoma cell line KYSE150,and to explore its molecular mechanism.Methods: 1.MTT assay,DAPI staining,cell adhesion assay,cell-extracellular matrix adhesion assay and scratch test were employed to detect the effect of the cell proliferation,apoptosis and anoikis,adhesion and migration ability of the cell lines which treated by different concentration(0??,0.5??,1??)of VX-680.2.Real-time PCR were used to detect the change of adhesion molecule CD44 and matrix metalloproteinase MMP-2 expression.3.Western blot wae used to detect the change of the apoptosis molecule Caspase3?PARP,adhesion molecule E-cadherin?CD44 and matrix metalloproteinase MMP-2 expression of the KYSE150 cell lines which treated different concentrations of VX-680.4.Western blot was used to explore the effects of different concentrations of VX-680 on the expression of p-ERK and p-AKT.Results:1.After esophageal cancer cell line KYSE150 treated by different concentrations of VX-680,With the increasing VX-680 concentration: MTT experimental results showed that the rate of cell proliferation decreased(P<0.05);DAPI staining showed that cell nuclear condensation and nuclear fragmentation were increased(P<0.001);Slow aggregation assay showed the cell particles were become bigger and tighter(P<0.001).Cell dissociation assay showed that cells were more difficiult separated than control groups(P<0.001);cell-matrix adhesion showed that the adhesion ability of the cells to three different extracellular matrix was gradually weakened(P<0.05);The results showed that the healing ability of the cells decreased gradually(P<0.001).2.Real-time PCR results showed that the expression of CD44 and MMP-2 decreased with the increase of VX-680 concentration.3.Westernblot showed that apoptosis Procaspase3 zymogen protein gradually decreased,PARP molecules appear obvious shear band;adhesion molecule expression of E-cadherin protein gradually increased and the expression of CD44 protein decreased;matrix metalloproteinase expression of MMP-2 protein decreased gradually with the increase of VX-680 concentration 4.Westernblot results showed that the expression of p-ERK and p-AKT decreased with the increase of VX-680 concentration.Conclusions: 1.The proliferation,migration,cell-matrix adhesion ability of esophageal carcinoma cells were significatly inhibited and the apoptosis?anoikis and cell-cell adhesion ability were enhanced after the treatment of different concentrations of Aurora kinase VX-680,the actiation of Caspase3,PARP and expression of E-cadherin,CD44,MMP-2,all those effects presented a dose-dependent manner.2.VX-680 may alter the biological behavior of KYSE150 cells in esophageal cancer through AKT and ERK pathway.