title: The antiangiogenic activity of a soluble fragment of the VEGFR extracellular domain.
abstract: Vascular endothelial growth factor (VEGF) is a key regulator of pathological angiogenesis and vascular permeability and overexpressed by most solid tumors. VEGF receptor-2 (VEGFR-2 or kinase-insert domain-containing receptor as it is called in human, KDR) is a specific receptor of VEGF with a high binding affinity. A solube recombinant extracellular domain 1-3 of human VEGFR-2 (rKDR1-3) was expressed in Escherichia coli (E. Coli) and purified from the bacterial periplasmic extracts by immobilized metal affinity chromatography and anion exchange chromatography to inhibit the VEGF-induced angiogenesis. A surface plasmon resonance (SPR) technology was adopted to analyze the affinity and kinetics constant between rKDR1-3 and VEGF165. Under the given experimental conditions, the association rate constant Ka was 1.06×10(5)M(-1) S(-1), the dissociation rate Kd was 6.09×10(-3) S(-1), the dissociation constant KD was 5.74×10(-8)M. The effect of rKDR1-3 on VEGF-induced endothelial cell proliferation was studied using MTT assay, scratch-wound healing assay and chorioallantoic membrane (CAM) assay. The results showed that rKDR1-3 could inhibit neovascularization and serve as a useful drug candidate in research, diagnostics and therapy of cancer.
authors.affiliation: State Key Laboratory of Natural Medicines (China Pharmaceutical University), School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Haixin
authors.initials: H
authors.lastname: Li
authors.forename: Wanlu
authors.initials: W
authors.lastname: Cao
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Chen
authors.forename: Desmond Omane
authors.initials: DO
authors.lastname: Acheampong
authors.forename: Haizhen
authors.initials: H
authors.lastname: Jin
authors.forename: Daojuan
authors.initials: D
authors.lastname: Li
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.forename: Min
authors.initials: M
authors.lastname: Wang
mesh_terms: D020533:Angiogenesis Inhibitors / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D049033:Chorioallantoic Membrane / Q000187:drug effects; D004730:Endothelium, Vascular / Q000187:drug effects; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D012995:Solubility; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000737:chemistry*
chemical_list: D020533:Angiogenesis Inhibitors; D010446:Peptide Fragments; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Antiangiogenic activity; BSA; Bovine serum album; CAM; Chorioallantoic membrane; Dissociation; E. Coli; EC; EGF; ELISA; Endothelial cells; Enzyme linked immunosorbent assay; Epidermal growth factor; Equilibrium constant; Escherichia coli; FBS; FGF; Fetal bovine serum; Fibroblast growth factors; IFN-γ; Interferon-γ; K(a); K(d); KD; KDR; Kinase-insert domain-containing receptor (as it is called in human); Monoclonal antibody; Periplasmic expression; SPR; Surface plasmon resonance; TNF-α; The kinetic constant for association; Tumor necrosis factor-α; VEGF; VEGF receptor-2; VEGFR-2; VEGFR2; Vascular endothelial growth factor; mAB