Gastrin promotes human colon cancer cell growth via CCK-2 receptor-mediated cyclooxygenase-2 induction and prostaglandin E2 production

Br J Pharmacol. 2005 Feb;144(3):338-48. doi: 10.1038/sj.bjp.0706053.

Abstract

The present study investigates the effects of gastrin-17 on human colon cancer HT-29 cells to examine whether gastrin receptor (CCK-2), cyclooxygenase (COX-1, COX-2) isoforms and prostaglandin receptor pathways interact to control cell growth. Reverse transcription (RT)-polymerase chain reaction (PCR) analysis demonstrated that HT-29 cells are endowed with the naive expression of CCK-2 receptor (short splice variant), COX-1, COX-2 and prostaglandin EP(4) receptor, but not gastrin. Gastrin-17 significantly promoted cell growth and DNA synthesis. Both these stimulating effects were abolished by L-365,260 or GV150013 (CCK-2 receptor antagonists), but were unaffected by SC-560 (COX-1 inhibitor). L-745,337 (COX-2 inhibitor) or AH-23848B (EP(4) receptor antagonist) partly reversed gastrin-17-induced cell growth, while they fully antagonized the enhancing action on DNA synthesis. HT-29 cells responded to gastrin-17 with a significant increase in prostaglandin E(2) release. This enhancing effect was completely counteracted by L-365,260, GV150013 or L-745,337, while it was insensitive to cell incubation with SC-560. Exposure of HT-29 cells to gastrin-17 was followed by an increased phosphorylation of both extracellular regulated kinases (ERK-1/ERK-2) and Akt. Moreover, gastrin-17 enhanced the transcriptional activity of COX-2 gene promoter and stimulated COX-2 expression. These latter effects were antagonized by L-365,260 or GV150013, and could be blocked also by PD98059 (inhibitor of ERK-1/ERK-2 phosphorylation) or wortmannin (inhibitor of phosphatidylinositol 3-kinase). Analogously, gastrin-17-induced prostaglandin E(2) release was prevented by PD98059 or wortmannin. The present results suggest that (a) in human colon cancer cells endowed with CCK-2 receptors, gastrin-17 is able to enhance the transcriptional activity of COX-2 gene through the activation of ERK-1/ERK-2- and phosphatidylinositol 3-kinase/Akt-dependent pathways; (b) these stimulant actions lead to downstream increments of COX-2 expression, followed by prostaglandin E(2) production and EP(4) receptor activation; (c) the recruitment of COX-2/prostaglandin pathways contributes to the growth-promoting actions exerted by gastrin-17.

MeSH terms

  • Antimetabolites
  • Blotting, Western
  • Bromodeoxyuridine
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Colonic Neoplasms / pathology*
  • Cyclooxygenase 2
  • DNA / biosynthesis
  • DNA / genetics
  • Dinoprostone / biosynthesis*
  • Enzyme Induction / drug effects
  • Gastrins / pharmacology*
  • HT29 Cells
  • Humans
  • Isoenzymes / biosynthesis
  • Luciferases
  • Membrane Proteins
  • Phosphatidylinositol 3-Kinases / metabolism
  • Promoter Regions, Genetic / genetics
  • Prostaglandin-Endoperoxide Synthases / biosynthesis*
  • RNA / biosynthesis
  • RNA / isolation & purification
  • Receptor, Cholecystokinin B / antagonists & inhibitors
  • Receptor, Cholecystokinin B / drug effects*
  • Receptors, Prostaglandin / drug effects
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / drug effects

Substances

  • Antimetabolites
  • Gastrins
  • Isoenzymes
  • Membrane Proteins
  • Receptor, Cholecystokinin B
  • Receptors, Prostaglandin
  • RNA
  • DNA
  • Luciferases
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • Phosphatidylinositol 3-Kinases
  • Bromodeoxyuridine
  • Dinoprostone