【摘要】 目的研究抗KDR单体Ycom1D3抑制VEGF诱导脐静脉内皮细胞(HUVEC)增殖的体外生物学活性。方法采用FACS鉴定Ycom1D3与抗原结合特异性,采用免疫共沉淀测定Ycom1D3阻断VEGF165刺激KDR酪氨酸激酶受体磷酸化作用,并采用[3H]-Thymidine掺入法、内皮细胞损伤愈合试验和内皮细胞血管腔形成实验进一步确定Ycom1D3抑制VEGF165诱导内皮细胞增殖的中和活性。结果Ycom1D3不但能与HUVEC结合,而且能阻断由VEGF165刺激HUVEC表面KDR酪氨酸激酶受体磷酸化,进而显著抑制VEGF165诱导HUVEC增殖、迁移及体外三维胶原模型上内皮细胞毛细血管样结构的形成。结论Ycom1D3可以通过封闭KDR而抑制VEGF活性,在肿瘤及其它血管新生疾病治疗中具有潜在应用前景。更多还原

【Abstract】 Objective To study the biological activity of a monoclonal antibody (Ycom1D3) against extracellular immunoglobulin (Ig)-like domain Ⅲ of vascular endothelial growth factor receptor KDR. Methods The specificity of Ycom1D3 was analyzed by FACS. The blocking effects of Ycom1D3 on phosphorylation of KDR tyrosine kinase receptor was detected by immunoprecipitation. [3H]-thymidine incorporation assay, wound healing assay, and in vitro angiogenesis assay were used to detect the influence on vascular endothelial growth factor-induced mitogenesis of human umbilical vascular endothelial cells (HUVECs). Results Ycom1D3 could bind specifically to the cell-surface expressed KDR, and then blocked effectively VEGF/KDR interaction and inhibited VEGF-stimulated KDR activation in HUVECs. Furthermore, the antibody efficiently neutralized VEGF-induced mitogenesis of HUVECs. Conclusion Ycom1D3 has potential applications in the treatment of cancer and other diseases where pathological angiogenesis is involved.更多还原