Impact of the transfer of sFlt-1 gene fragments on the ERK1/2 pathway of VEGF in vitro.
OBJECTIVE
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.
Department of Ophthalmology, Shanghai No. 6 People's Hospital, Shanghai, PR China.
Dan
D
Jiang
Hongchen
H
Gu
Qiang
Q
Wu
Xiaoliang
X
Wang
Min
M
Zhang
Beiwen
B
Song
D015153:Blotting, Western; D002452:Cell Count; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D004574:Electromagnetic Fields; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D005786:Gene Expression Regulation / Q000502:physiology*; D005947:Glucose / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D000860:Hypoxia / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics*
D010446:Peptide Fragments; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D005947:Glucose