血管扩张刺激磷蛋白的磷酸化位点突变对PDGF-BB诱导细胞迁移的影响  

Effect of VASP mutant phosphorylation on migration of endothelial cells induced by PDGF-BB

作　　者：张亚辉[1,2] 张德玲[1] 欧阳静萍[1] 

机构地区：[1]武汉大学医学院病理生理学教研室,湖北武汉430071 [2]郧阳医学院病理生理学教研室,湖北十堰442000

出　　处：《中国病理生理杂志》2010年第1期32-36,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30470680)

摘　　要：目的:探讨血管扩张刺激磷蛋白(VASP)Ser157、Ser239位点的磷酸化对血小板源性生长因子BB(PDGF-BB)诱导细胞迁移的影响及其作用差异。方法:构建VASPSer157和Ser239位点突变的真核表达质粒pcDNA3.1(+)/VASP-S157A和pcDNA3.1(+)/VASP-S239A,分别转染ECV304细胞,并筛选出它们的稳定表达细胞株,通过Transwell小室跨膜迁移实验检测2种细胞株的迁移速度。结果:经PDGF-BB作用后,2种稳定表达细胞株的跨膜迁移数量明显少于对照组(P<0.05),而2种细胞株之间无显著差异(P>0.05)。结论:VASPSer157和Ser239磷酸化位点突变减弱了PDGF-BB诱导的细胞迁移,Ser157和Ser239磷酸化位点对VASP的功能均具有重要影响。AIM： To investigate the effect of vasodilator- stimulated phosphoprotein （VASP） phosphorylation on the cell migration induced by platelet - derived growth factor BB （ PDGF - BB ）, and to identify which phosphorylation site was more important among the three phosphorylation sites, namely Ser157, Ser239 and Thr278. METHODS： Two phosphorylation mutants of VASP, pcDNA3.1 （ ＋ ）/VASP- S157A and pcDNA3.1 （ ＋ ）/VASP- S239A, were constructed and respectively transfected into the cultured ECV304 cells by means of liposome. The stable expression cells were screened by using antibiotic G418. Protein expression of VASP was measured by Western blotting. The ECV304 cell migration was evaluated using Transwell chamber. RESULTS： Compared to control group, the cell migration was significantly inhibited in ECV304 transfected with VASP - S157A and VASP -S239A （ P 〈 0. 05 ）, although slight differences existed between VASP- S157A and VASP-S239A transfected cells （P 〉0. 05）. CONCLUSION： VASP mutation on the phosphorylation sites of Ser157 and Ser239 inhibits cell migration, and the phosphorylation sites of Ser157 and Ser239 both greatly affect the function of VASP.

关 键 词：血管扩张刺激磷蛋白 细胞运动 血小板源性生长因子 

分 类 号：R363.1[医药卫生—病理学]