Vascular endothelial growth factor-regulated gene expression in endothelial cells: KDR-mediated induction of Egr3 and the related nuclear receptors Nur77, Nurr1, and Nor1.
OBJECTIVE
The program of gene expression regulated by vascular endothelial growth factor (VEGF) remains poorly understood. The aim of this study was to identify VEGF-regulated genes in human umbilical vein endothelial cells.


RESULTS
VEGF-regulated gene expression was analyzed by screening Affymetrix oligonucleotide arrays and quantitative, real-time, reverse transcription-polymerase chain reaction. The most strongly induced genes were the NR4A nuclear receptor family members Nur77, Nurr1, and Nor1 and the zinc-finger transcription factor Egr3. VEGF also induced rapid expression of Down syndrome candidate region 1, cyclooxygenase-2, tissue factor, stanniocalcin-1, the serine/threonine kinase Cot, and eps15 homology domain-containing protein. VEGF-induced NR4A family and Egr3 expression was blocked by a KDR inhibitor, and placental growth factor and basic fibroblast growth factor weakly increased expression of these genes. Induction of NR4A genes was mediated via intracellular Ca2+, protein kinase C- and calcineurin-dependent pathways. VEGF increased protein expression of Nurr1 and Nur77 and decreased Nur77 phosphorylation at the negative regulatory site serine 351.


CONCLUSIONS
VEGF induces expression of NR4A nuclear receptors and Egr3 via KDR and KDR-mediated signaling mechanisms. The genes identified here are novel candidates as key early mediators of VEGF-induced endothelial functions.
Liu
Dan
D
BHF Laboratories, Department of Medicine, University College London, London, UK.
Jia
Haiyan
H
Holmes
David Ian Roderick
DI
Stannard
Anita
A
Zachary
Ian
I
D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D051776:Early Growth Response Protein 3; D004730:Endothelium, Vascular / Q000502:physiology*; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D026901:Membrane Transport Proteins; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism*; D014470:Umbilical Cord / Q000098:blood supply; D042461:Vascular Endothelial Growth Factor A / Q000502:physiology*; D014680:Veins
D004268:DNA-Binding Proteins; C079945:EGR3 protein, human; D026901:Membrane Transport Proteins; C539821:NR4A1 protein, human; C539882:NR4A2 protein, human; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; C479458:OSCP1 protein, human; D011506:Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; D051776:Early Growth Response Protein 3