粘着斑激酶在血管内皮细胞黏附和迁移中的作用  

Role of focal adhesion kinases in adhesion and migration of endothelial cells

作　　者：沈阳[1] 沈楠[1] 刘文辉[1] 赖怡[1] 李良[1] 康敏[1] 付强[1] 刘肖珩[1,2] 

机构地区：[1]四川大学华西基础医学与法医学院,生物医学工程研究室,成都610041 [2]西藏大学医学院,拉萨850002

出　　处：《医用生物力学》2011年第2期128-132,共5页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(10972148);中国博士后科学基金面上资助项目(20090461340);第三批中国博士后基金特别资助项目(201003702)

摘　　要：目的采用粘着斑激酶(focal adhesion kinases,FAK)抑制剂抑制FAK在Y397位点的酪氨酸磷酸化,测定不同浓度的FAK抑制剂的对内皮细胞黏附、迁移及下游信号Rac1蛋白表达的影响,探索粘着斑激酶在内皮细胞黏附和迁移中的作用。方法运用内皮损伤模型(划痕法)测定FAK抑制剂在24、8、、24 h各时间点对EA.hy 926细胞迁移的影响。Western blot结合免疫荧光测定不同浓度的0～250 nmol/mL的FAK抑制剂的加入对Rac1蛋白分布和表达的影响。结果随着FAK抑制剂浓度的增加,细胞迁移距离减少,Rac1蛋白表达逐渐减弱。结论抑制FAK的磷酸化将抑制内皮细胞的黏附和迁移的生物学行为,下游Rac1蛋白表达降低。内皮细胞的黏附和迁移与FAK-Rho GTPases信号轴相关。Objective To investigate the effect of focal adhesion kinases（FAK） inhibitor with different concentration on the adhesion and migration of endothelial cells（ECs） and the expression of downstream Rac1 protein,and to explore the role of FAK in adhesion and migration of ECs by using FAK inhibitor to inhibit the phosphoryla-tion of Y397 site of FAK.Method Scratch wound migration assay was performed to examine the effect of FAK inhibitor with different concentration（from 0 nmol/mL to 250 nmol/mL） on ECs migration at 2,4,8 and 24 h,respectively.Western blot combined with immunofluorescence analysis were performed to determine the effect of FAK inhibitor with different concentration on distribution and expression of Rac1 protein.Results With the concentration of FAK inhibitor increased,ECs migration distance and the Rac1 protein expression decreased.Conclusions The inhibition of FAK phosphorylation could inhibit cell adhesion and migration with the decrease in downstream Rac1 protein,and ECs adhesion/migration was related to FAK-Rho GTPases signaling pathways.

关 键 词：粘着斑激酶 内皮细胞 蛋白表达 细胞黏附 细胞迁移 

分 类 号：R3[医药卫生—基础医学]