反义FAK寡核苷酸抑制FAK-ERK1/2介导的平滑肌细胞迁移和粘附  

FAK ANTISENSE OLIGODEOXYNUCLEOTIDES INHIBIT VASCULAR SMOOTH MUSCLE CELL MIGRATION AND ADHESION MEDIATED BY FAK-ERK1/2 SIGNALING PATHWAY

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血管病研究所,北京100034

出　　处：《药学学报》2002年第5期334-338,共5页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目 (3 973 0 2 2 0 )

摘　　要：目的　研究FAK ERK1 2信号通路在平滑肌细胞迁移和粘附中的作用及FAK反义寡核苷酸对其的调控作用。方法　通过纤粘连蛋白 (fibronectin ,FN)诱导平滑肌细胞迁移和粘附 ,以免疫沉淀和Westernblot方法检测粘着斑激酶 (focaladhesionkinase,FAK)和细胞外调控激酶 1 2 (extracellularregulationkinase ,ERK1 2 )及其磷酸化的表达量。将FAK反义寡核苷酸 (antisenseoligodeoxynucleotides,ODNs)经脂质体转染细胞 ,观察转染后反义ODN对FAK和ERK磷酸化、细胞粘附和迁移的影响。结果　FN在有效诱导平滑肌细胞迁移和粘附时FAK和ERK1 2也呈明显表达 ,FN2 0 μg·mL- 1 可使磷酸化处于较高表达量。脂质体可有效地介导ODNs转染。转染后FAK及ERK1 2磷酸化表达量明显减少。结论　由活化的FAK和ERK1 2介导的信号转导促进了细胞外基质诱导的SMCs迁移和增殖 ,反义FAKODNs可有效地对此进行抑制。AIM To study the effects of FAK ERK1/2 signaling pathway and FAK antisense oligodeoxynucleotides (ODNs) on vascular smooth muscle cell (SMC) migration and adhesion stimulated by fibronectin (FN). METHODS Migration and adhesion of cultured SMCs were stimulated by different concentrations of FN, FAK, ERK1/2. And their phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense ODNs were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation, SMCs migration and adhesion were also measured by modifing Boyden Chamber and morphological enumeration, respectively. RESULTS FAK were expressed when SMCs adhesion and migration were successfully simulated by FN (5, 10, 20, 40, 60 μg·mL -1 ), high contents of FAK and ERK1/2 phosphorylation were detected by 20 μg·mL -1 FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid. FAK and ERK1/2 phosphorylation were inhibited magnificently after FAK antisense ODNs transfection. Cell migration stimulated by FN 10, 20, 40 and 60 μg·mL -1 were reduced by 23 26%, 21 63%, 19 31% and 17 88% respectively ( P <0 05). SMCs adhesive spreading in 5～60 μg·mL -1 FN groups were reduced by 17 89%～27 67% ( P <0 05). CONCLUSION FAK ERK1/2 mediated signal transduction play important roles in SMCs migration and adhesion stimulated by extracellular matrix. The process can be inhibited by FAK antisense ODNs effectively.

关 键 词：粘着斑激酶 细胞外调控激酶1/2 反义寡核苷酸 平滑肌细胞 

分 类 号：R963[医药卫生—微生物与生化药学] R329.28[医药卫生—药理学]