title: Recombinant human vascular endothelial growth factor receptor 1 effectively inhibits angiogenesis in vivo.
abstract: Vascular endothelial growth factor (VEGF) plays an important role in both physiological and pathological angiogenesis. VEGF receptor‑1 (VEGFR‑1) acts as a decoy VEGF receptor that enables the regulation of VEGF on the vascular endothelium. In the present study, the recombinant human VEGFR1D1‑3/Fc (rhVEGFR‑1), which contains key domains for VEGF binding, was cloned and expressed in Chinese hamster ovary (CHO) cells. The rhVEGFR‑1 protein was purified using protein‑A affinity chromatography. The molecular weight of rhVEGFR‑1 was found to be ~162 and 81 kD in non‑reducing and reducing SDS‑PAGE, respectively. The majority of the final protein products were in the dimeric conformation. Western blot analysis revealed that rhVEGFR‑1 was only capable of binding to the full glycan form of rhVEGF‑165 and rhVEGF‑121. The dissociation constant for the binding of rhVEGFR‑1 to VEGF‑165, detected using Biacore, was 285 pM. In addition, rhVEGFR‑1 inhibited the proliferation and migration of human microvascular endothelial cells. In vivo experiments also demonstrated that rhVEGFR‑1 inhibited chicken chorioallantoic membrane neovascularization and angiogenesis in nude mice. In conclusion, an anti‑angiogenic recombinant soluble VEGFR was expressed (up to 5 mg/l) in CHO cells and was shown to be capable of inhibiting neovascularization in vivo and in vitro.
authors.affiliation: Department of General Oncology, Chinese PLA General Hospital, Beijing 100853, P.R. China.
authors.forename: Jinliang
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Minglei
authors.initials: M
authors.lastname: Shi
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Yongyi
authors.initials: Y
authors.lastname: Xi
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Guanyi
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing 100071, P.R. China.
authors.forename: Xianwen
authors.initials: X
authors.lastname: Hu
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D016466:CHO Cells; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D003412:Cricetulus; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011993:Recombinant Fusion Proteins; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000494:pharmacology*
chemical_list: D020533:Angiogenesis Inhibitors; D011993:Recombinant Fusion Proteins; D040281:Vascular Endothelial Growth Factor Receptor-1