The Research On The Role And The Mechanism Of Rho/ ROCK Signaling Pathway In Invasiveness Of U87 Human Glioblastma Cell In Vitro

Objective To investigate the effect of Rho/Rho-associated coiled coil-forming kinase (ROCK) signaling pathway on the proliferation,migration and metastasis of U87 human glioblastma cell(U87MG),a malignant human glioma cell line.And explore the mechanism of Rho/ ROCK signaling pathway。Methods The malignant human glioma cell line (U87MG) was cultivated in vitro. U87cells were treated with 0, 5, 10, 25, 50and 100μmol/L concentrations Y-27632 for 12h, 24h in vitro and the inhibition rates were examined by MTT assay. After U87 cells were incubated with final concentrations of 0, 10, 25 and 50μmol/L Y-27632 for 12h, investigated the alteration of ROCK activity,invasiveness and movement of cells,by Rho-kinase Assay Kit,transwell test and motility test.U87 cells grew 12h with different concentrations (0, 10, 25 and 50μmol/L) Y-27632 in vitro and then the expression of MMP-2 and MMP-9 proteins were confirmed using Western Blotting.Results MTT assay showed thatY-27632 has not demonstrated effect on proliferation of U87 cells. In transwell and motility test the ability of invasiveness and movement of cells was deeply inhibited.The concertrition was higher,the inhibition is stronger.After action of different concentration of Y-27632 in U87cells,ROCK activity of cells correspondingly decreased.The concentration was higher,the activity is lower. Westorn blot results indicated that under the action of different concentration of Y-27632,total MMP-2 and MMP-9 amount were decreasing,obviously in MMP-2.Conclusion In vitro experiments the ability of invasiveness and movement of U87 cells was deeply inhibited by blocked Rho/ ROCK signaling pathway,and the expression of MMP-2 and MMP-9 proteins were decreasing.The Rho/ROCK signaling pathway was very important to invasiveness and movement of U87 cells .