Induction of MUC5AC mucin by conjugated bile acids in the esophagus involves the phosphatidylinositol 3-kinase/protein kinase C/activator protein-1 pathway

Cancer. 2011 Jun 1;117(11):2386-97. doi: 10.1002/cncr.25796. Epub 2010 Dec 14.

Abstract

Background: Bile reflux contributes to the development of esophageal injury and neoplasia. The mucin 5AC (MUC5AC) is absent in the normal squamous epithelium of the esophagus but is strongly expressed in Barrett esophagus (BE). The objective of this study was to determine whether and how bile acids influence the expression of MUC5AC in the esophagus.

Methods: MUC5AC expression was studied by immunohistochemistry and immunoblotting in human tissues, in tissues from a rat model of BE, and in SKGT-4 cultured esophageal epithelial cells. MUC5AC transcription was studied by real-time polymerase chain reaction and transient transfection assays.

Results: MUC5AC was absent from normal squamous epithelium but was present in 100% of Barrett specimens and in 61.5% of human esophageal adenocarcinoma tissues that were examined. MUC5AC protein expression was induced to a greater degree by conjugated bile acids than by unconjugated bile acids, and this occurred at the transcriptional level. In the rat reflux model, MUC5AC mucin was expressed abundantly in tissues of BE stimulated by duodenoesophageal reflux. Conjugated bile acids induced AKT phosphorylation in SKGT-4 cells but had no effect on extracellular signal-regulated protein kinases 1 and 2, c-Jun N-terminal kinase, or protein-38 kinase phosphorylation. The phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 and a dominant-negative protein kinase C (AKT) construct prevented the induction of MUC5AC by conjugated bile acids. Transactivation of AP-1 by conjugated bile acids coincided with MUC5AC induction, and cotransfection with a dominant-negative activator protein-1 (AP-1) vector decreased MUC5AC transcription and its induction.

Conclusions: Conjugated bile acids in the bile refluxate contribute to MUC5AC induction in the esophagus. This occurs at the level of transcription and involves activation of the PI3K/AKT/AP-1 pathway.

Keywords: Barrett esophagus; MUC5AC mucin, bile reflux; activator protein-1; phosphatidylinositol 3-kinase pathway.

Publication types

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

MeSH terms

  • Adenocarcinoma / genetics
  • Adenocarcinoma / metabolism
  • Animals
  • Barrett Esophagus / genetics
  • Barrett Esophagus / metabolism
  • Bile Acids and Salts / metabolism*
  • Bile Acids and Salts / pharmacology
  • Bile Reflux / genetics
  • Bile Reflux / metabolism
  • Disease Models, Animal
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Esophageal Neoplasms / genetics
  • Esophageal Neoplasms / metabolism
  • Esophagus / metabolism*
  • Gene Expression
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Male
  • Mucin 5AC / genetics
  • Mucin 5AC / metabolism*
  • Mucous Membrane / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Protein Kinase C / metabolism*
  • Rats
  • Signal Transduction*
  • Transcription Factor AP-1 / metabolism*
  • Transcription, Genetic

Substances

  • Bile Acids and Salts
  • Mucin 5AC
  • Transcription Factor AP-1
  • Phosphatidylinositol 3-Kinases
  • Protein Kinase C