Impact of the Transfer of sFlt-1 Gene Fragments on the ERK1/2 Pathway of VEGF In Vitro

Abstract

Purpose: To investigate whether various sFlt-1 gene fragments affect the biological functions and ERK1/2 pathway of vascular endothelial growth factor (VEGF) under conditions of hypoxia or in the presence of high glucose concentrations in vitro. Methods: Plasmids expressing loops 2–3 and loops 2–4 of sFlt-1 were packed in carboxymethylated dextran-coated nanoparticles and transferred into human umbilical vein endothelial cells (HUVECs), which were then cultured under hypoxia or in a high-glucose environment. The proliferation and migration of HUVECs were examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and low-power microscopy, respectively. Western blot analyses were performed to detect p-ERK1/2 protein expression. Results: After transfection with the sFlt-1(2-3) or sFlt-1(2-4) gene fragment, the proliferation and migration of HUVECs were markedly reduced, and p-ERK1/2 protein expression was down-regulated under both hypoxic and high-glucose conditions. The impacts on the proliferation, migration of HUVECs, and on p-ERK1/2 protein expression did not differ significantly between the sFlt-1(2-3) and sFlt-1(2-4) gene fragments. Conclusions: Both sFlt-1(2-3) and sFlt-1(2-4) gene fragments inhibited the proliferation and migration of HUVECs, as well as signal transduction in the ERK1/2 pathway of VEGF.