血管内皮生长因子受体2胞外区3(KDR3)血管抑制活性研究  

Antiangiogenic Activity Research of the Vascular Endothelial Growth Factor Receptor 2 Extracellular Region 3(KDR3)

作　　者：曹婉璐[1] 张娟[1] 涂孝洁[1] 许卓斌[1] 金海珍[1] 王旻[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2013年第3期203-206,共4页Pharmaceutical Biotechnology

基　　金：国家自然科学基金资助项目(NSFC81072561;NSFC81102364;NSFC81273425);国家大学生创新性实验计划项目(G12071)

摘　　要：血管内皮生长因子(VEGF)是一种重要的血管生成调节因子。血管内皮生长因子受体2(VEGFR2,人VEGFR2也称为Kinase insert domain-containing receptor,KDR)是VEGF的特定膜受体之一。血管内皮生长因子阻断剂可以通过阻断VEGF-VEGFR2信号通路来调节重要的抑癌过程。目前尚没有研究VEGFR2胞外3区(KDR3)抑制血管活性的文献。因此,该研究旨在用大肠杆菌表达可溶性KDR3,并研究其抑制血管生成的活性。采用表面等离子体共振(SPR)技术检测到可溶性表达KDR3与VEGF16 5之间的平衡解离常数为54 nmol/L。采用划痕损伤修复实验来证明KDR3能够显著抑制EA.hy926细胞的迁移能力。最后通过鸡胚尿囊膜实验表明,KDR3在体内同样能够抑制新生血管生成。因此,该文初步论证外源性KDR3作为一种血管内皮生长因子阻滞剂的能力。Vascular endothelial growth factor （VEGF） is a vital mediator of angiogenesis. VEGF receptor 2 （VEGFR2） interacts with VEGF,as one of its specific membrane receptors. Researchers have found out that a VEGF blocker can restrain some vital processes of tumor suppression by blocking the signal pathway of VEGF-VEGFR2. However, there is no literature evidence of investi- gation on the antiangiogenic ability of single domain 3 of VEGFR-2 （ KDR3 ）, as the key domain in the signal transduction of VEGF. In this article, Escherichia coli Rosetta-gami （DE3）/pET22b was taken to express the soluble KDR3. The binding between KDR3 and VEGF165 was tested with BiacoreX100 （GE）. Scratch-wound healing assay was used to evaluate KDR3＇s cell migration ability. The in vivo anti-angiogenic effect of KDR3 was tested with chicken embryo chorioallantoic membrane （CAM） model. The outcome showed that the soluble KDR3 was expressed successfully with prokaryotic expression system. The surface plasmon resonance （SPR） tech- nology was adopted to test the binding between KDR3 and VEGF165, which equihbrium dissociation rate constant （Kd） was 54 nmol/L. KDR3 can suppress EA. hy926 cell＇ migration with a dose-dependent tendency. Also the in vivo investigation showed that KDR3 is capable of inhibiting neovascularization, adopting the CAM assay. This article has therefore provided a novel antiangiogenic drug candidate relating to VEGF-VEGFR2 pathway.

关 键 词：血管内皮生长因子受体 血管抑制能力 鸡胚尿囊膜模型 表面等离子共振技术 划痕修复实验 血管阻断剂 

分 类 号：Q51[生物学—生物化学]