粘着斑激酶活性对气道上皮细胞粘附迁移的影响  

Effects of Focal Adhesion Kinase Phosphorylation on Adhesion and Migration of Human Airway Epithelial Cells Stimulated by Fibronectin

作　　者：许丽[1] 汪涛[1] 张珍祥[1] 徐永健[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院呼吸疾病研究室,武汉430030

出　　处：《华中科技大学学报（医学版）》2005年第3期297-300,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30370623)

摘　　要：目的研究粘着斑激酶(FAK)磷酸化对细胞外基质成份诱导的气道上皮细胞粘附、迁移的影响,探讨FAK活化对气道上皮细胞损伤后修复的影响机制。方法通过纤维连接蛋白(FN)诱导培养的气道上皮细胞,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Westernblot和免疫共沉淀检测FAK表达水平及其酪氨酸磷酸化程度;以反义FAK寡核苷酸(ODNs)经脂质体转染细胞,观察其对FAK表达和磷酸化、细胞粘附和迁移的影响。结果FN诱导的气道上皮细胞粘附率和迁移速度明显增高,同时FAK表达水平及其酪氨酸磷酸化程度显著增高;经脂质体转染了反义FAK寡核苷酸的气道上皮细胞粘附率和迁移速度明显降低,同时FAK表达水平及其酪氨酸磷酸化程度降低,且与气道上皮细胞粘附率和迁移速度的降低成显著正相关。结论FAK是细胞外基质诱导气道上皮细胞粘附、迁移的重要信号分子,其活化促进气道上皮细胞的粘附和迁移,在气道上皮细胞损伤后修复过程中起重要作用。Objective To study the effects of focal adhesion kinase (FAK) phosphorylation on the cell adhesion and migration of human airway epithelial cell stimulated by fibronectin.Methods Cultured human airway epithelial cells were stimulated by fibronectin. The rate of adhesion was measured by counting the adhered airway epithelial cells; The velocity of cell migration was measured by wound-repair process; The FAK expression level was detected by Western blot and the degree of tyrosine phosphorylation by immunoprecipitation. FAK antisense oligodeoxynucleotides (ODNs) were transfected into human airway epithelial cells by actionic lipid method.Results The rate of adhesion and the velocity of cell migration of human airway epithelial cells stimulated by fibronectin was significantly increased. The expression level and phosphorylation of FAK was increased obviously. FAK antisense ODNs could decrease the cell adhesion and inhibit the velocity of cell migration, meanwhile the expression and phosphorylation of FAK were inhibited drastically. The latter changes were negatively correlated to the rate of adhesion and the velocity of cell migration.Conclusion FAK phosphorylation and FAK-mediated signal transduction play important roles in adhesion and migration of human airway epithelial cells, so activation of FAK may be involved in the wound-repair process of airway epithelia.

关 键 词：粘着斑激酶 寡核苷酸 反义 气道上皮细胞 

分 类 号：R332.2[医药卫生—人体生理学]