Vascular endothelial growth factor induction by prostaglandin E2 in human airway smooth muscle cells is mediated by E prostanoid EP2/EP4 receptors and SP-1 transcription factor binding sites

J Biol Chem. 2005 Aug 26;280(34):29993-30000. doi: 10.1074/jbc.M414530200. Epub 2005 Jun 21.


Prostaglandin E2 (PGE2) can increase endothelial vascular endogrowth factor A (VEGF-A) production but the mechanisms involved are unclear. Here we characterized the transcriptional mechanisms involved in human airway smooth muscle cells (HASMC). PGE2 increased VEGF-A mRNA and protein but not mRNA stability. PGE2 stimulated the activity of a transiently transfected 2068-bp (-2018 to +50) VEGF-A promoter-driven luciferase construct. Functional 5' deletional analysis mapped the PGE2 response element to the 135-bp sequence (-85/+50) of the human VEGF-A promoter. PGE2-induced luciferase activity was reduced in cells transfected with a 135-bp VEGF promoter fragment containing mutated Sp-1 binding sites but not in cells transfected with a construct containing mutated EGR-1 binding sites. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay confirmed binding of Sp-1 to the VEGF promoter. PGE2 increased phosphorylation of Sp-1 and luciferase activity of a transfected Sp-1 reporter construct. PGE receptor agonists EP2 (ONO-AE1 259) and EP4 (ONO-AE1 329) mimicked the effect of PGE2, and reverse transcription-PCR, Western blotting, and flow cytometry confirmed the presence of EP2 and EP4 receptors. VEGF protein release and Sp-1 reporter activity were increased by forskolin and isoproterenol, which increase cytosolic cAMP, and the cAMP analogue, 8-bromoadenosine-3',5'-cyclophosphoric acid. These studies suggest that PGE2 increases VEGF transcriptionally and involves the Sp-1 binding site via a cAMP-dependent mechanism involving EP2 and EP4 receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Cell Survival
  • Cells, Cultured
  • Chromatin Immunoprecipitation
  • Colforsin / pharmacology
  • Cyclic AMP / metabolism
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Dinoprostone / metabolism*
  • Dose-Response Relationship, Drug
  • Gene Deletion
  • Humans
  • Isoproterenol / pharmacology
  • Luciferases / metabolism
  • Molecular Sequence Data
  • Mutation
  • Myocytes, Smooth Muscle / cytology*
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA / metabolism
  • RNA, Messenger / metabolism
  • Receptors, Prostaglandin E / metabolism*
  • Receptors, Prostaglandin E, EP2 Subtype
  • Receptors, Prostaglandin E, EP4 Subtype
  • Sp1 Transcription Factor / metabolism*
  • Time Factors
  • Trachea / cytology
  • Transcription, Genetic
  • Transfection
  • Vascular Endothelial Growth Factor A / biosynthesis*
  • Vascular Endothelial Growth Factor A / metabolism


  • PTGER2 protein, human
  • PTGER4 protein, human
  • RNA, Messenger
  • Receptors, Prostaglandin E
  • Receptors, Prostaglandin E, EP2 Subtype
  • Receptors, Prostaglandin E, EP4 Subtype
  • Sp1 Transcription Factor
  • Vascular Endothelial Growth Factor A
  • Colforsin
  • RNA
  • Cyclic AMP
  • Luciferases
  • Cyclic AMP-Dependent Protein Kinases
  • Dinoprostone
  • Isoproterenol