Peng Xu, Ueda Hiroki, Zhou Hongming, Stokol Tracy, Shen Tang-Long, Alcaraz Ana, Nagy Tamas, Vassalli Jean-Dominique, Guan Jun-Lin
Department of Molecular Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Cardiovasc Res. 2004 Dec 1;64(3):421-30. doi: 10.1016/j.cardiores.2004.07.012.
Focal adhesion kinase is implicated in the regulation of cell adhesion, migration, survival, and cell-cycle progression. However, the functions of focal adhesion kinase in endothelial cell (EC) in vivo remain unclear. This study aims to examine the role of FAK in EC function and angiogenesis in vivo by transgenic mice approach.
We generated transgenic mice which overexpressed chicken FAK in vascular endothelial cell under the control of the Tie-2 promoter and enhancer. FAK transgene was detected by RT-PCR, immunoprecipitation, and Western blot. The effect of FAK overexpression on angiogenesis was determined using skin wound healing and ischemia skeleton muscle models.
Expression of FAK transgene was detected in all vessel-rich tissues. Expression of FAK protein was verified by antibody specific for the exogenous chicken FAK in lung homogenates and isolated EC. In the wound-induced angiogenesis model, the number of vessels in the granulation tissue of healing wound was significantly increased in the transgenic mouse compared to that of wild-type control mice. Similarly, in the ischemia skeleton muscle model, the density of capillaries was significantly increased in the transgenic mouse.
These results indicate that FAK may play an important role in the promotion of angiogenesis in vivo.
粘着斑激酶参与细胞粘附、迁移、存活及细胞周期进程的调控。然而,粘着斑激酶在体内内皮细胞(EC)中的功能仍不清楚。本研究旨在通过转基因小鼠方法研究FAK在体内EC功能及血管生成中的作用。
我们构建了在Tie-2启动子和增强子控制下在血管内皮细胞中过表达鸡FAK的转基因小鼠。通过RT-PCR、免疫沉淀和蛋白质印迹检测FAK转基因。使用皮肤伤口愈合和缺血骨骼肌模型确定FAK过表达对血管生成的影响。
在所有富含血管的组织中均检测到FAK转基因的表达。通过针对外源性鸡FAK的抗体在肺匀浆和分离的EC中验证了FAK蛋白的表达。在伤口诱导的血管生成模型中,与野生型对照小鼠相比,转基因小鼠愈合伤口肉芽组织中的血管数量显著增加。同样,在缺血骨骼肌模型中,转基因小鼠的毛细血管密度显著增加。
这些结果表明FAK可能在体内促进血管生成中起重要作用。