Effects Of MEK/ERK Inhibitor U0126 On Proliferation,Migration And Invasion Of Human Tongue Squamous Cell Carcinoma 8113 Cells

Objective:To investigate the effects of U0126 on Proliferation,migration and invasion of human tongue squamous cell carcinoma 8113 cells,and explore its mechanisms.Method:The Tca8113 cells were cultured in vitro and treated by different concentration of U0126(25?50?100 ?mol/L),a MEK/ERK inhibitor.The effect of U0126 on the proliferation of Tca8113 cells were assessed by MTT assay.The migration and invasion of Tca8113 cells were measured by wound healing assay and Transwell assay in vitro respectively.The Protein locations and levels of ERK1/2,p-ERK1/2,p-MEKK1,MEKK1 were detected by immunofluorescence and Western blot method;the protein levels of EMT-related protein E-cadherin Vimentin were detected by Western blot method.Results:1.Compared with control group,the proliferation of Tca8113 cells were obviously inhibit by treated with 25?50 and 100?mol/LU0126 for 24?48 and 72h(P<0.01and P<0.05);and compared with 25?mol/L group,inhibited by 50 and 100?mol/L of U0126 for 24?48 and 72h were significantly(P<0.01 and P<0.05);and compared with 50?mol/L group,by treated 100?mol/L of U0126 significantly inhibit the proliferation of Tca8113 cells(P<0.01),which showed concentration dependent manner.Compared with 24h,the proliferation of Tca8113 cells were obviously inhibit by treated with 25?50 and 100?mol/L U0126 for 48 and 72h(P<0.01);and Compared with 48h,the proliferation were significantly inhibited by treated for 72h(P<0.01),which showed time dependent manner.2.Compared with control group,The migrating distance of Tca8113 cells were markedly inhibited after treated with 25.50 and 100?mol/LU0126 for 24?48 and 72h(P<0.01).3.Compared with control group,The number of Tca8113 cells that invaded the underside of the porous polycarbonate membrane were significantly decreased by treated with 25?50?100?mol/L of U0126 in the below chamber for 48h,and the relative invasion rate were79%?51%?35%respectively(P<0.01),which showed concentration dependent manner.4.The Immunofluorescence analysis showed that the expression of MEKK1 mainly located in the cytoplasm,the p-MEKK1 and ERK1/2mainly located in the cytoplasm and nuclear membrane,and the p-ERK1/2 located in the nuclear membrane and nucleus?The fluorescence density of p-ERK1/2 were decreased with 25?50?100?mol/L of U0126 for 48h,which showed concentration dependent manner.whereas the ERK1/2?p-MEKK1?MEKK1 has no significant difference.5.The western blot analysis revealed that the expression of p-ERK1/2 about Tca8113 cells were decreased than control group after treated with 25?50?100?mol/L of U0126 for 48h(P<0.01),whereas the expression of ERK1/2?p-MEKK1?MEKK1 protein has no significant difference.6.The western blot analysis revealed that compared with control group,the expression of Vimentin about Tca8113 cells were decreased after treated with 25?50?100?mol/L of U0126 for 48h(P<0.01),whereas the E-cadherin were increased(P<0.01),and the ERK1/2?p-MEKK1?MEKK1 protein has no significant difference.Conclusion:1.U0126 significantly inhibit the proliferation?migration and invasion of Tca8113 cell.2.Its mechanism may be associated with inhibiting the phosphorylation of ERK1/2 by MAPK/ERK1/2 signaling pathways.3.Its mechanism may be related to increase the expression of E-Cadherin and decrease the expression of Vimentin,and then to inhibit Epithelial-mesenchymal transition.ERK1/2 signal transduction pathway will become a potential therapeutic target for cancer.