Osthole Inhibits Proliferation,Invasion And Metastasis Of Vulvar Squamous Carcinoma Cells By PI3K/Akt Pathway

ObjectiveTo investigate the effect of osthole on the proliferation and invasion and metastasis of vulvar squamous cell carcinoma cell line SW962,and to explore its effect on cell epithelial mesenchysis and possible signaling pathways.MethodsThe vulvar squamous cell carcinoma cell line SW962 was cultured in vitro.The morphology of cells was observed by crystal violet staining.The effects of osthole on apoptosis and cell proliferation of SW962 cells were observed by MTT assay,DAPI staining and flow cytometry.Scratch test,colony test and Transwell test were used to detect the effect of osthole on SW962 invasion and metastasis.Western blot was used to detect apoptosis,cell cycle,invasion and metastasis and expression of epithelial-like proteins.Its possible signal path.ResultsOsthole could inhibit the proliferation of SW962 cells and inhibit the formation of colonies in a concentration-dependent manner.Blocking the cell cycle in G1 phase,up-regulate the expression of cell cycle-associated protein p21,and down-regulate the expression of cyclin D1 and CDK4(P<0.05).Osthol could induce apoptosis in a concentration-dependentmanner,and the expression of proapoptotic protein Bax and cleaved caspase-3 is significantly increased,while the expression of anti-apoptotic protein Bcl-2 is significantly decreased(P<0.05),and the expression of PI3 K and p-Akt protein is significantly decreased expression(P<0.05).Osthole inhibited cell invasion and metastasis and was positively correlated with the concentration,which up-regulated the expression of EMT-related protein E-cadherin,and down-regulated the expression of ?-catenin,wnt1,MMP2 and Vimentin(P<0.05).When IGF-1 was used to activate PI3K/Akt pathway,the expression of Bcl-2,?-catenin,Wnt1 and Vimentin was increased,and the expression of E-cadherin was decreased(P<0.05).Osthole could reverse the effect of IGF.After nucleoplasmic separation,IGF-1 was found to increase the expression of ?-catenin in the nucleus(P<0.05),and activate the nucleation of ?-catenin.Osthole could antagonize this nucleation(P<0.05).ConclusionOsthole can inhibit the proliferation of vulvar squamous cell carcinoma SW962 cells in a time-and dose-dependent manner,promote its apoptosis,arrest cell cycle in G1 phase,and inhibit cell invasion and metastasis in a concentration-dependent manner.It can inhibit the wnt/?-catenin pathway by inhibiting the PI3K/Akt pathway and block the nuclear import of ?-catenin,promote apoptosis and inhibit epithelial mesenchymal transition.