U0126对人舌鳞癌Tca8113细胞增殖、迁移及侵袭的影响

Effects of U0126 on proliferation,migration and invasion of human tongue squamous cell carcinoma Tca8113 cells

[目的]探讨U0126对人舌鳞癌Tca8113细胞增殖、迁移及侵袭的影响.[方法]用不同浓度MEK/ERK抑制剂U0126(25,50,100μmol/L)处理Tca8113细胞24,48,72 h,采用MTT法检测细胞增殖抑制情况,划痕实验检测细胞迁移距离,Transwell实验检测48 h时的细胞侵袭率,细胞免疫荧光染色和Western blot法检测48h时的细胞ERK1/2,p-ERK1/2,MEKK1,p-MEKK1在细胞内定位和蛋白表达情况及E-cadherin和Vimentin表达水平.[结果]U0126对细胞的增殖、迁移和侵袭具有明显的抑制作用(P<0.01),且呈现出浓度和时间依赖性;细胞中磷酸化的ERK1/2主要表达在细胞核膜和核内,U0126对磷酸化的ERK1/2表达具有明显的抑制作用(P<0.01);与对照组比较,U0126各剂量组E-cadherin蛋白表达水平均明显升高(P<0.01),而Vimentin蛋白表达水平明显降低(P<0.01),且均呈现出浓度依赖性.[结论]U0126可抑制Tca8113细胞的增殖、迁移和侵袭,其机制认为可能与抑制ERK1/2活化、促进E-cadherin表达及抑制Vimentin表达有关系.

OBJECTIVE To investigate the effects of U0126 on proliferation,migration and invasion of human tongue squamous cell carcinoma Tca8113 cells.METHODS The Tca8113 cells were treated by different concentration of MEK/ERK inhibitor U0126（25,50,100μmol/L）for 24,48 and 72 hours.MTT assay was used to detect the inhibition of cell proliferation,and the migration distance was detected by wound healing assay.Transwell assay was used to detect the cell invasion at 48 hour.Immunofluorescence staining and Western blot method were used to detect the protein locations and levels of ERK1/2,p-ERK 1/2,MEKK1 and p-MEKK1 at 48 hour,and detect the expressions of E-cadherin and Vimentin.RESULTS U0126 had obvious inhibitory effects on cell proliferation,migration and invasion（P〈0.01）,and showed a concentration-dependent and time-dependent manner.P-ERK1/2 was mainly expressed in the nuclear membranes and nuclei of cells,and U0126 significantly inhibited the expression of p-ERK1/2（P〈0.01）.As compared with the control group,the expression of E-cadherin in each U0126 groups was significantly increased（P〈0.01）,while the expression of Vimentin was obviously decreased（P〈0.01）,all of them showed concentration dependence.CONCLUSION U0126 could inhibit the proliferation,migration and invasion of Tca8113 cells.The mechanism might be related to the inhibition of ERK1/2 activation,promote the expression of Ecadherin and inhibit the expression of Vimentin.