title: COX-2 induction by unconjugated bile acids involves reactive oxygen species-mediated signalling pathways in Barrett's oesophagus and oesophageal adenocarcinoma.
abstract: OBJECTIVE
  Bile reflux contributes to oesophageal injury and neoplasia. COX-2 is involved in both inflammation and carcinogenesis; however, the precise mechanisms by which bile acids promote COX-2 expression in the oesophagus are largely unknown. We analysed the molecular mechanisms that govern bile acid-mediated expression of COX-2 in Barrett's oesophagus and oesophageal adenocarcinoma (OA).
  
  
  METHODS
  The effects of bile acids on COX-2 expression were analysed in immortalised Barrett's oesophagus and OA cells using immunoblotting and transient transfections. Pharmacological inhibitors, phospho-specific antibodies, dominant-negative mutants and siRNA techniques were used to identify relevant signalling pathways. Flow cytometry and reactive oxygen species (ROS) scavengers were used to examine ROS involvement. Immunohistochemistry was performed on oesophageal mucosa obtained from an established rat model of bile reflux.
  
  
  RESULTS
  Unconjugated bile acids potently stimulated COX-2 expression and induced AKT and ERK1/2 phosphorylation in concert with COX-2 induction. These findings were mimicked in the in vivo rat model. Dominant-negative (DN) AKT and LY294002 (PI3K inhibitor) or U0126 (MEK-1/2 inhibitor) blocked chenodeoxycholic acid (CD) and deoxycholic acid (DC) mediated COX-2 induction. CD and DC also induced CREB phosphorylation and AP-1 activity. CREB-specific siRNA and DN AP-1 blocked CD and DC-induced COX-2 induction. Finally, CD and DC increased intracellular ROS, while ROS scavengers blocked COX-2 induction and the signalling pathways involved.
  
  
  CONCLUSIONS
  Unconjugated bile acids induce CREB and AP-1-dependent COX-2 expression in Barrett's oesophagus and OA through ROS-mediated activation of PI3K/AKT and ERK1/2. This study enhances our understanding of the molecular mechanisms by which bile acids promote the development of oesophageal adenocarcinoma.
authors.affiliation: Department of Gastrointestinal Medicine and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030-4009, USA.
authors.forename: Shumei
authors.initials: S
authors.lastname: Song
authors.forename: Sushovan
authors.initials: S
authors.lastname: Guha
authors.forename: Kaifeng
authors.initials: K
authors.lastname: Liu
authors.forename: Navtej S
authors.initials: NS
authors.lastname: Buttar
authors.forename: Robert S
authors.initials: RS
authors.lastname: Bresalier
mesh_terms: D000230:Adenocarcinoma / Q000209:etiology / Q000378:metabolism* / Q000517:prevention & control; D000818:Animals; D001471:Barrett Esophagus / Q000209:etiology / Q000378:metabolism*; D001647:Bile Acids and Salts / Q000494:pharmacology*; D050882:CREB-Binding Protein / Q000378:metabolism; D002450:Cell Communication / Q000502:physiology; D002471:Cell Transformation, Neoplastic; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D004938:Esophageal Neoplasms / Q000209:etiology / Q000378:metabolism*; D005764:Gastroesophageal Reflux / Q000150:complications*; D006801:Humans; D007150:Immunohistochemistry; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D001647:Bile Acids and Salts; D017382:Reactive Oxygen Species; D051546:Cyclooxygenase 2; D050882:CREB-Binding Protein; D048052:Mitogen-Activated Protein Kinase 3