Gao Min, Liu Xiaoheng, Sun Heng, Ren Hongyi, Wang Lijuan, Shen Yang
Institute of Biomedical Engineering, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2013 Jun;30(3):567-71.
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 inhibitor with 50nmol/mL on ECs migration at 0, 5, 10, 30, 60 and 120min, respectively. And immunofluorescence analysis 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 results 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 increasing 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.
血管内皮细胞迁移诱导的肿瘤血管生成是肿瘤生长和转移的必要条件。本研究旨在探讨粘着斑激酶（FAK）抑制剂（50nmol/mL）对内皮细胞（ECs）粘附、迁移以及粘着斑蛋白纽蛋白、踝蛋白和桩蛋白表达的影响。进行划痕伤口迁移试验，分别检测50nmol/mL的FAK抑制剂在0、5、10、30、60和120分钟时对ECs迁移的影响。并进行免疫荧光分析，以检测用FAK抑制剂处理2小时内ECs中F-肌动蛋白的表达。采用蛋白质免疫印迹法检测FAK抑制剂对纽蛋白、踝蛋白和桩蛋白表达的影响。结果显示，与对照组相比，用FAK抑制剂处理的ECs迁移距离和F-肌动蛋白表达显著降低，随着FAK抑制剂处理时间的增加，纽蛋白水平无显著差异。然而，踝蛋白和桩蛋白在5-10分钟呈相同下降趋势，但在30分钟时缓慢上升，随后又下降。本研究结果证明，阻断FAK磷酸化可通过下调粘着斑蛋白抑制血管内皮细胞粘附和迁移，从而可能抑制肿瘤血管生成。这可能为肿瘤治疗提供一种新方法。
