Ang II and EGF synergistically induce COX-2 expression via CREB in intestinal epithelial cells

J Cell Physiol. 2008 Jan;214(1):96-109. doi: 10.1002/jcp.21167.


Cyclooxygenase (COX)-2 derived prostaglandins (PGs) play a major role in intestinal inflammation and colorectal carcinogenesis. Because COX-2 is the rate-limiting step in the production of PGs, mechanisms that regulate COX-2 expression control PG production in the cell. Using the non-tumorigenic, rat intestinal epithelial cell, IEC-18, we demonstrate that co-activation of endogenously expressed AT(1) receptor and EGFR resulted in synergistic expression of COX-2 mRNA and protein involving transcriptional and post-transcriptional mechanisms. Ang II and EGF induced transient phosphorylation of ERK, p38(MAPK) and CREB. Co-stimulation with Ang II and EGF prolonged phosphorylation of ERK, p38(MAPK), and CREB. The p38(MAPK) selective inhibitor, SB202190, but not the MEK selective inhibitor, PD98059, or the EGFR kinase inhibitor, AG1478, inhibited Ang II-dependent COX-2 expression and CREB phosphorylation. EGF-dependent COX-2 expression and CREB phosphorylation were inhibited by SB202190, PD98059, and AG1478. Inhibition of CREB expression using two separate RNAi methods blocked COX-2 expression by Ang II and EGF. Expression of a dominant negative CREB mutant inhibited Ang II- and EGF-dependent induction of the COX-2 promoter. Ang II induced luciferase expression in cells transfected with the CRE-luc reporter vector and cells co-transfected with Gal4-luc reporter vector and a Gal4-CREB expression vector. Chromatin immunoprecipitation assays demonstrated CREB binding to the proximal rat COX-2 promoter region containing a CRE cis-acting element. These results indicate that co-stimulation with Ang II and EGF synergistically induced COX-2 expression in these intestinal epithelial cells through p38(MAPK) mediated signaling cascades that converge onto CREB.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Angiotensin II / pharmacology*
  • Animals
  • Cells, Cultured
  • Chromatin Immunoprecipitation
  • Culture Media, Serum-Free
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Cyclooxygenase 2 / metabolism*
  • Dinoprostone / analysis
  • Dinoprostone / metabolism
  • Drug Synergism
  • Electrophoretic Mobility Shift Assay
  • Epidermal Growth Factor / pharmacology*
  • Epithelial Cells / drug effects*
  • Epithelial Cells / enzymology
  • Epithelial Cells / metabolism
  • Epoprostenol / analysis
  • Epoprostenol / metabolism
  • Gene Expression Regulation, Enzymologic / drug effects*
  • Intestines / cytology*
  • Luciferases / analysis
  • Luciferases / metabolism
  • Models, Biological
  • RNA Interference
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Rats
  • Transfection


  • Culture Media, Serum-Free
  • Cyclic AMP Response Element-Binding Protein
  • RNA, Messenger
  • RNA, Small Interfering
  • Angiotensin II
  • Epidermal Growth Factor
  • Epoprostenol
  • Luciferases
  • Cyclooxygenase 2
  • Dinoprostone