Activated focal adhesion kinase involved in adhesion and migration of vascular smooth muscle cells stimulated by fibronectin  

粘着斑激酶活化对平滑肌细胞粘附和迁移的影响

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

机构地区：[1]北京大学第一医院心脏科,北京,100034

出　　处：《Chinese Medical Journal》2002年第4期494-497,共4页中华医学杂志（英文版）

摘　　要：OBJECTIVE: To study the effects of focal adhesion kinase (FAK) phosphorylation on smooth muscle cells (SMCs) adhesion and migration stimulated by fibronectin. METHODS: Adhesion and migration of cultured SMCs were stimulated by different concentrations of fibronectin (FN), FAK and its phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. SMCs adhesion and migration were also measured by morphological enumeration and modified Boyden Chambers, respectively. RESULTS: FAK were expressed when SMCs adhesion and migration were successfully simulated by different concentrations of FN. FAK phosphorylation were detected only at 20 microg/ml FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid and FAK phosphorylation was inhibited substantially. The SMCs migration rate in the 5 - 60 microg/ml FN groups was reduced by 17.89% - 27.67%. Cell migration stimulated by FN at 10, 20, 40 and 60 microg/ml were reduced by 23.26%, 21.63%, 19.31% and 17.88%, respectively (P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(fibronectin, FN)诱导培养的平滑肌细胞粘附迁移,以免疫沉淀和Western blot方法检测粘着斑激酶(focal adhesion kinase, FAK)及其磷酸化的表达量.将FAK反义寡核苷酸(antisense oligodeoxynucleotides, 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,20?μg/ml FN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,5-60?μg/ml不同浓度FN组,细胞铺展率减少17.89%-27.67%,10、 20、 4 0和60?μg/ml FN组迁移细胞数也分别显著减少23.26%、21.63%、19.31%、17.88%(P <0.05).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,反义FAK ODNs可有效地对此进行抑制.

关 键 词：Animals Cell Adhesion Cell Movement Cells  Cultured DNA  Antisense Dose-Response Relationship  Drug Enzyme Activation FIBRONECTINS Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Muscle  Smooth  Vascular PHOSPHORYLATION Protein-Tyrosine Kinase Rats TRANSFECTION 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]