目的:探讨表皮生长因子受体突变体Ⅲ(epidermal growth factor receptor mutation typeⅢ,EGFRvⅢ)促进入胶质瘤细胞迁移运动的可能机制。方法:利用反转录病毒感染技术建立EGFRvⅢ过表达的人胶质瘤细胞U87 EGFRvⅢ,琼脂滴细胞迁移实验和Transwell细胞迁移实验观察EGFRvⅢ对细胞迁移运动的影响;Western印迹法检测黏着斑激酶(focal adhesion kinase,FAK)磷酸化的变化。将FAK突变体FAK(Y397F)导入U87 EGFRvⅢ细胞,观察细胞侵袭运动变化。结果:EGFRvⅢ促进入胶质瘤细胞U87的迁移运动,同时使FAK397位点磷酸化水平上调。将FAK突变体Y397F转入U87 EGFRvⅢ细胞,FAK磷酸化水平下调同时细胞迁移能力下降。结论:EGFRvⅢ通过上调FAK397位点磷酸化水平来促进胶质瘤细胞的迁移。
Objective: To investigate the mechanism underlying the promoting effects of epidermal growth factor receptor mutation type Ⅲ ( EGFRv Ⅲ ) on the migration of glioma cells. Methods: Human glioma cell line U87 EGFRv Ⅲ was established by retroviral infection of the parental glioma cells. The effect of EGFRv Ⅲ on the migration of the cells was detected by gel-drop assay and Transwell migration assay. The phosphorylation of FAK was detected by Western blotting. The FAK Y397F mutant was transfected into U87EGFRv Ⅲ cells and the change in cell migration was observed. Results: EGFRv Ⅲ promoted the migration of human glioma U87 cells and increased the FAK phosphorylation level at Tyr397. Transfection with FAK Y397F mutant reduced FAK phosphorylation level in glioma cells U87EGFRv Ⅲ and induced the decrease in the migration abilities of the cells. Conclusion: EGFRv Ⅲ promoted the migration of glioma cells by up-regulating FAK phosphorylation levels.