Abstract:
Background: Blocking vascular endothelial
growth factor receptor (VEGFR) is a successful
approach for inhibiting vascular endothelial
growth factor (VEGF) signaling. Small molecules
impairing the interaction of VEGF with VEGF
receptors have been synthesized and evaluated
in this research.
Methods: In this study, we amplified and
cloned the cDNA of VEGFR fragments. After
expression of the fragments in Escherichia
coli (E. coli), they were purified by immobilized
metal affinity chromatography (IMAC).
The biological activity of recombinant KDR
fragments was evaluated by human umbilical
vein endothelial cells (HUVEC) proliferation
assay.
Results: The sodium dodecyl sulfate
polyacrylamide gel electrophoresis (SDS-PSGE)
and immune-blotting results confirmed the
production and purification of recombinant
VEGFR fragments. The purified VEGFR2-III
domain, VEGFR2-II-III domains, and VEGFR1-II
domain showed 24%, 36%, and 27%
inhibition effect in HUVEC proliferation assay,
respectively. The recombinant VEGFR2-IIIII
domains strongly inhibited formation of
capillary-like structures (CLS).
Conclusions: The produced recombinant
proteins will serve as small soluble molecules
with an inhibitory effect against VEGF in antiangiogenesis
researches.
