粘着斑激酶活化对平滑肌细胞粘附和迁移的影响  

Effects of FAK phosphorylation on adhesion and migration of smooth muscle cells stimulated by fibronectin

作　　者：尹航[1] 汪丽蕙[1] 彭旭[1] 霍勇[1] 夏春芳[1] 唐朝枢[1] 

机构地区：[1]北京医科大学第一医院心血管研究所,北京100034

出　　处：《中国病理生理杂志》2002年第6期593-596,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目 (No .39730 2 2 0 )

摘　　要：目的 :研究粘着斑激酶磷酸化在细胞外基质成份诱导平滑肌细胞粘附和迁移中的作用。方法 :通过纤粘连蛋白 (FN)诱导培养的平滑肌细胞粘附迁移 ,以免疫沉淀和Westernblolt方法检测粘着斑激酶 (FAK)及其磷酸化的表达量。将FAK反义寡核苷酸 (ODNs)经脂质体转染细胞 ,观察对FAK磷酸化、细胞粘附铺展和迁移的影响。结果 :FN在显著诱导平滑肌细胞粘附和迁移时 ,FAK也呈明显表达 ,2 0mg/LFN可使其磷酸化处于较高表达量。脂质体可有效地介导ODNs转染 ,转染效率为 (86 7± 4 5 ) % ,FAK磷酸化表达量明显减少 ,5 - 6 0mg/L不同浓度FN组 ,细胞铺展率减少 17 89% - 2 7 6 7% ,10、2 0、4 0和 6 0mg/LFN组迁移细胞数也分别显著少 2 3 2 6 %、2 1 6 3%、19 31%、17 88% (P <0 0 5 )。结论 :活化的FAK是细胞外基质诱导SMCs粘附和迁移的重要信号分子 ,由其介导的信号转导促进了这一过程 。AIM: To study the effect of focal adhesion kinase (FAK) phosphorylation on the adhesion and migration of smooth muscle cells (SMCs) stimulated by fibronectin (FN). METHODS: Cultured SMCs were stimulated by different concentrations of FN.FAK expression and phosphorylation were detected by immunoprecipitation and Western blot. To investigate the modulating effect of FAK on tyrosine phosphorylation and SMCs adhesion and migration, FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by actionic lipid method. RESULTS: FAK expression increased when SMCs adhesion and migration were induced by FN at different concentrations used in the experiment. However, FAK phosphorylation was only observed by FN stimulation at concentration of 20 mg/L. FAK antisense ODNs inhibited FAK phosphorylation magnificently. The migration rates of SMCs were reduced by 17.89%-27.67% when FN was used at concentration from 5 mg/L to 60 mg/L. The decreased migrating cell numbers were showed the same patterns. The apoptotic SMCs were 33.57% higher than that control ( P <0 05). CONCLUSION: FAK phosphorylation and FAK-mediated signal transduction play important roles in SMCs adhesion and migration. The process can be inhibited by FKA antisense ODNs effectively.

关 键 词：粘着斑激酶 细胞粘附 纤粘连蛋白 平滑肌细胞 细胞迁移 血管成形术 血管再狭窄 

分 类 号：R654.306[医药卫生—外科学]