Effects Of Aurora Kinase Inhibitor VX-680 On Apoptosis,Migration And Angiogenesis In Human Hepatocellular Carcinoma HepG2 Cells

Objective:1.To investigate the effects of Aurora protein kinase inhibitor VX-680 on the proliferation,apoptosis,adhesion,migration and tubule formation of HepG2 cells.2.To investigate the molecular mechanism of VX-680 suppressing the malignant phenotype of HepG2 cells.Methods:1.MTT assay,plate clone formation assay,DAPI staining,cell-cell adhesion assay,cell-extracellular matrix adhesion assay,wound healing assay and tubular formation assay were used to observe the effect of different concentrations(0 ?mol/L,3.125 ?mol/L,6.25 ?mol/L,12.5 ?mol/L)VX-680 on the proliferation,apoptosis,adhesion,migration and tubular formation of human hepatoma HepG2 cells.2.Western blot was used to detect the change of the apoptosis molecules(Caspase-3 and PARP),adhesion molecules(E-cadherin,CD44)and metastasis-related molecular MMP-2,vascular endothelial growth factor A(VEGFA)expression.3.Western blot was used to detect the effects of different concentrations of VX-680 on the ERK and AKT signaling pathways in HepG2 cells.Results:1.With the increasing VX-680 concentration,the results of MTT assay and plate clone formation showed that the proliferation rate of cells decreased gradually(P<0.05).The results of DAPI staining showed the cell nucleus concentration and nucleus fragmentation ratio gradually increased(P<0.001).The results of cell-cell adhesion experiments showed that the aggregates formed by the cells became larger and larger(P<0.001),and the cells were more and more difficult to be dispersed(P<0.001).The cell-extracellular matrix adhesion assay showed that the adhesion ability of the cells to the three extracellular matrix gradually weakened(P<0.05).Wound healing assay showed that the healing ability of cells in the experimental group was gradually weakened and healed time prolonged(P<0.001);Tubule formation experiments showed that the number of tubules decreased gradually(P<0.001).2.Western blot results showed that with the increase of VX-680 concentration,the expression level of Caspase-3 proenzyme and PARP decreased,the cleavage bands of PARP protein increased,E-cadherin expression increased,the expression of CD44,MMP-2 and VEGFA decreased gradually.3.The results of Western blot showed that the expression of p-ERK and p-AKT protein decreased gradually with the increase of VX-680 concentration,while the total protein ERK and AKT had not changed significantly.Conclusions:1.Aurora protein kinase inhibitor VX-680 can inhibit HepG2 cell proliferation in a concentration and time dependent manner,promote cell apoptosis,enhance cell-cell homogeneity adhesion force,weaken cell-extracellular matrix adhesion and migration ability,and inhibit HUVEC tubule formation.2.VX-680 can promote the apoptosis of hepatoma cells by regulating the activity of Caspase-3 and PARP.VX-680 can increase the homogeneous adhesion of cell-cell adhesion,reduce cell-extracellular matrix adhesiveness and inhibit migration by increasing the expression of E-cadherin and decreasing expression of CD44 and MMP-2.VX-680 can reduce the angiogenic ability of HUVEC cells by down-regulating the expression of VEGFA in HepG2 cells.3.VX-680 may inhibit the malignant phenotype of HepG2 cells by inhibiting the AKT and ERK signaling pathways.