FAK通过粘着斑蛋白调控血管内皮细胞迁移的研究  

Study on FAK Regulation of Migration of Vascular Endothelial Cells Depending upon Focal Adhesion Proteins

作　　者：高敏[1] 刘肖珩[1] 孙亨[1] 任弘毅[1] 王利娟[1] 沈阳[1] 

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实验室,成都610041

出　　处：《生物医学工程学杂志》2013年第3期567-571,共5页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(10972148;11172198);国家基础科学人才培养基金能力提高资助项目(J1103604)

摘　　要：血管内皮细胞(VECs)迁移导致的肿瘤内血管新生是肿瘤生长和转移的必要条件。本研究目的在于探索粘着斑激酶(FAK)引起的粘着斑蛋白变化以及对VECs黏附和迁移的影响。采用粘着斑激酶抑制剂(50nmol/mL)抑制FAK在Y397位点的磷酸化,划痕法测定FAK抑制剂在0、5、10、30、60、120min对VECs迁移的影响,免疫荧光测定FAK抑制剂作用120min后细胞骨架的变化,Western blot测定FAK抑制剂的加入对粘着斑vinculin、talin和paxillin蛋白表达的变化情况。结果表明,FAK抑制剂加入后,与对照组相比细胞迁移距离明显减少,胞间连丝被打断,vinculin蛋白表达在各时间点基本一致,而talin和paxillin表达在初始的5～10min出现降低趋势,30min有缓慢升高,30min后表达再次降低。本研究结果证明了阻断FAK的磷酸化可下调几种粘着斑蛋白复合物的表达,在细胞水平上抑制了VECs的黏附和迁移行为。该研究结果为由VECs迁移引起的肿瘤内血管新生的治疗提供了一定的实验基础。Tumor angiogenesis induced by vascular endothelial cells （VECs） migration is a necessary condition for tumor growth and metastasis. The purpose of this study is to investigate the effect of focal adhesion kinase （FAK） inhibitor （50nmol/mL） on the adhesion and migration of endothelial cells（ECs） and the expression of focal adhesion proteins vinculin, talin and paxillin. Scratch wound migration assay was performed to examine the effect of FAK in- hibitor with 50nmol/mL on ECs migration at 0, 5, 10, 30, 60 and 120rain, respectively. And immunofluorescence a- nalysis was performed to detect the expression of F-actin in ECs treated with FAK inhibitor within 2h. Western blot was carried out to determine the effect of FAK inhibitor on expression of vinculin,talin and paxillin proteins. The re- sults showed that the migration distance and the expression of F-actin in ECs treated with FAK inhibitor decreased significantly compared with that of the controls, and the level of vinculin showed no significant difference with in- creasing of treated time of FAK inhibitor. However, the talin and paxillin showed an identical decreasing tendency in 5-10min, but slowly going up in 30min and then after subsequently decreasing. The results of this study proved that blocking phosphorylation of FAK could inhibit VECs adhesion and migration by downregulating focal adhesion proteins so that it may inhibit tumor angiogenesis. This may provide a new approach for tumor therapy.

关 键 词：粘着斑激酶 粘着斑蛋白 血管内皮细胞 细胞黏附 细胞迁移 

分 类 号：R730.2[医药卫生—肿瘤]