摘要
目的研究FIP200(FAK-family interacting protein of 200 kDa)对细胞外基质成份诱导的呼吸道上皮细胞粘附和迁移的影响及其作用机制。方法通过纤连接蛋白(finbronectin,FN)诱导培养呼吸道上皮细胞的粘附迁移,以FIP200反义寡核苷酸(ODN)经脂质体转染细胞;以计数法测定细胞粘附率,以损伤实验测定细胞迁移速度,以Western blot检测FIP200和FAK蛋白表达水平;以免疫共沉淀检测FIP200和FAK结合情况和FAK磷酸化程度。结果与40 mg/L FN组相比,经脂质体转染了FIP200反义寡核苷酸的气道上皮细胞粘附率和迁移速度明显增高,FIP200表达水平显著降低,FAK表达水平无明显变化,与FIP200结合的FAK显著降低,FAK酪氨酸磷酸化程度明显增高。结论内源性FIP200和FAK的结合抑制FAK的活化,从而抑制呼吸道上皮细胞的粘附和迁移,内源性FIP200作为FAK的抑制剂而存在;FN能够促使FAK和FIP200结合的解离而活化FAK,从而促进细胞粘附和迁移。
Objective To investigate the effects of FIP200 on the cell adhesion and migration of human airway epithelial cell stimulated by fibronectin. Methods The human airway epithelial cells were cultured and stimulated by fibronectin. FIP200 antisense oligodeoxynucleotides（ODNs） were transfected into human airway epithelial cells by actionic lipid method. The rate of adhesion was measured by counting the adhered airway epithelial cells and the velocity of cell migration was measured by wound-repair experiment; FIP200 and FAK expression level was analyzed by western blot;The association of FIP200 with FAK and the degree of Tyrosine phosphorylation of FAK by immunoprecipitation. Results FIP200 antisense ODNs improved the adhesion and migration of human airway epithelial cells; Meanwhile the expression of FIP200 was inhibited drastically,the expression of FAK was normal, but the association of FIP200 with FAK decreased significantly and the phosphorylation of FAK were increased drastically. Conclusion The association of endogenous FIP200 with FAK can inhibit the activation of FAK, by which FIP200 can inhibit cell adhesion and migration of human airway epithelial cells,so we can say endogenous FIP200 function as a protein inhibitor for FAK kinase activity.
出处
《江西医学院学报》
2006年第3期23-26,共4页
Acta Academiae Medicinae Jiangxi
