VASP磷酸化对PDGF-BB诱导细胞迁移的影响  

Role of VASP and PKA in Cell Migration Induced by PDGFBB

作　　者：别婧华[1] 欧阳静萍[1] 张德玲[1] 

机构地区：[1]武汉大学医学院病理生理学教研室,武汉430071

出　　处：《武汉大学学报（医学版）》2006年第6期687-690,715,共5页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:30470680)

摘　　要：目的:探讨血管扩张刺激磷蛋白(VASP)对血小板源性生长因子B(PDGF-BB)诱导的细胞迁移的影响及其机制。方法:通过细胞划痕损伤实验检测ECV304细胞迁移速度,用非放射性酶法检测该细胞中蛋白激酶A(PKA)的活性,用Western blot检测细胞中磷酸化VASP(P-VASP)表达的变化。结果:经PDGF-BB作用后,ECV304细胞向划痕内迁移的距离较对照组明显增加(P<0.05),胞内PKA活性明显增加(P<0.05),细胞内磷酸化VASP的表达显著增强(P<0.05);而预先给予PKA抑制剂H89处理能显著地抑制PDGF-BB介导的ECV304的迁移效应(P<0.01),明显降低PDGF引起的胞内PKA活性升高(P<0.01),抑制PDGF-BB对VASP的磷酸化作用(P<0.05)。结论:PDGF-BB通过PKA信号传导通路,使其下游底物VASP磷酸化,磷酸化的VASP介导细胞移动。Objective. To investigate the effect of vasodilator-stimulated phosphoprotein （VASP） on the cell migration induced by platelet-derived growth factor （PDGF）. Methods, The cell migration velocity of ECV304 cell line was detected by cell scratch wound assays. Activity of cyclic AMPdependent protein kinase（PKA） within the cells was detected by PepTag assay for non-radioactive detection. Also, the expression of P-VASP was detected by Western blot assay. Results. The cell scratch wound assays and the Western blot assay demonstrated that the distance of the cell migration of the experimental group treated with PDGF-BB was greater than that of the control group （P〈0.05）, the activity of PKA within cells obviously increased （P〈0.05）, and the expression of phosphorylated-VASP significantly strengthened after 30 minutes＇ stimulation of PDGF-BB on the cells （P〈0.05）. At the same time, when the cells were pretreated with H89, the inhibitor of PKA, the migration effect of ECV304 cells mediated by PDGF-BB was remarkably inhibited （P〈 0.01）, the activity of PKA within cells which increased formerly by PDGF was apparently inhibited （P〈0.01）, and the phosphorylated effect of PDGF-BB on the VASP was also inhibited（P〈 0.05）. Conclusion. VASP, a substrate of PKA, has been specially activated by stimulating PKA signal conduction pathway, and then phosphorylated-VASP played a role in cell migration.

关 键 词：血管扩张刺激磷蛋白 蛋白激酶A 血小板源性生长因子B 细胞迁移 ECV304 

分 类 号：R363.1[医药卫生—病理学]