Impacts of aucubin on viability and epithelial mesenchymal transformation of glioblastoma cells by regulating RhoA/ROCK signaling pathway
[Objective]To investigate the impacts of aucubin(AU)on the viability and epithelial-mesenchymal transition(EMT)of glioblastoma(GBM)cells,and to explore its mechanism of action.[Methods]U87 were grouped into control group,low concentration group,medium concentration group,high concentration group,Y-27632 group,and high concentration+Y-27632 group.Cell counting kit-8(CCK-8)method was applied to detect cell viability;flow cytometry was applied to detect cell apoptosis;Transwell cell experiment was applied to detect cell migration and invasion;Western blot was applied to detect the expression of matrix metalloproteinase(MMP)2,MMP9,Vimentin,E-cadherin,N-cadherin,RAS homologous gene family member A(RhoA),Rho-related coiled protein kinase(ROCK)1,ROCK2.The nude mouse model of GBM was constructed,and was grouped into nude mice control group,AU group,Y-27632 group,and AU+Y-27632 group;the mass and volume of tumors were measured,immunohistochemical method was applied to detect the expression of RhoA,ROCK1,and ROCK2 proteins in transplanted tumor tissue.[Results]The viability of GBM cells gradually decreased with the increase of AU concentration(P＜0.05);U87 was selected as the follow-up experimental cells,and 10,25,50 μmol/L was selected as the follow-up experimental concentration of AU.Compared with the control group,the cell viability,migration and invasion cell counts,MMP2,MMP9,N-cadherin,Vimentin,RhoA,ROCK1,and ROCK2 expression in the low,medium,and high concentration AU groups and Y-27632 group were obviously decreased,the apoptosis rate and E-cadherin expression were obviously increased(P＜0.05);the changes in high concentration AU and Y-27632 group were more obvious(P＜0.05).Both AU and Y-27632 were able to inhibit the mass and volume of transplanted tumors,and reduce the expression of RhoA/ROCK signaling pathway proteins(P＜0.05).[Conclusion]AU can inhibit the viability,migration,invasion and EMT of GBM cells and promote cell apoptosis,which may be related to the inhibition of RhoA/ROCK signaling pathway.
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