论文部分内容阅读
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。
Objective To study the in vitro biological activity of anti-KDR monomer Ycom1D3 in inhibiting the proliferation of human umbilical vein endothelial cells (HUVECs) induced by VEGF. Methods The specificity of Ycom1D3 binding to antigen was identified by FACS. Ycom1D3 was used to block the phosphorylation of KDR tyrosine kinase receptor by blocking VEGF165 by immunoprecipitation. [3H] -Thymidine incorporation method, endothelial cell injury healing assay and endothelial The experiment of cell lumen formation further confirmed that Ycom1D3 inhibits VEGF165-induced neutralizing activity of endothelial cells. Results Ycom1D3 could not only bind to HUVEC but also block the phosphorylation of KDR tyrosine kinase receptor on HUVECs stimulated by VEGF165, and then significantly inhibit the VEGF165-induced proliferation and migration of HUVECs and the formation of endothelial capillary capillary-like structures in three-dimensional collagen model . Conclusion Ycom1D3 can inhibit the activity of VEGF by blocking KDR and has potential application in the treatment of tumors and other angiogenesis diseases.