title: Upregulation of miRNA-143, -145, -192, and -194 in esophageal epithelial cells upon acidic bile salt stimulation.
abstract: Barrett's esophagus (BE) is a metaplastic condition of the distal esophagus that occurs because of chronic gastroesophageal reflux. Previous studies have identified BE-specific microRNAs (miRNAs) in comparison with normal squamous epithelium (SQ). We hypothesized that BE-specific miRNAs could be induced in esophageal SQ cells by exposure to acid and/or bile salts. We aimed to determine whether BE-specific miRNAs are upregulated in an esophageal SQ cell line (Het-1A) in an environment with acid and/or bile salts and whether this is nuclear factor-κB (NF-κB) dependent. Acid and/or bile salt incubations were performed in Het-1A cells. Experiments were performed with or without inhibiting the NF-κB pathway. Quantitative reverse transcriptase polymerase chain reaction was performed to determine expression of miRNA-143, -145, -192, -194, cyclo-oxygenase-2 (COX2), mucin 2 (MUC2), and sex determining region Y-box 9. For validation, we determined levels of these miRNAs in biopsies from patients with reflux esophagitis and normal SQ. Significantly increased expression levels of miRNA-143 (2.7-fold), -145 (2.6-fold), -192 (2.0-fold), -194 (2.2-fold), COX2, MUC2, and sex determining region Y-box 9 were found upon acidic bile salt incubation, but not upon acid or bile salt alone. NF-κB pathway inhibition significantly decreased miRNA-143, -192, -194, COX2, and MUC2 expression. Additionally, miRNA-143, -145 and -194 expression was increased in reflux esophagitis biopsies compared with normal SQ, but no changes were found in miRNA-192 expression. Our findings suggest that upregulation of BE-specific miRNAs by acidic bile may be an early event in the transition of SQ to BE and that their expression is partly regulated by the NF-κB pathway.
authors.affiliation: Department of Gastroenterology and Hepatology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Bus
authors.forename: P D
authors.initials: PD
authors.lastname: Siersema
authors.forename: R E
authors.initials: RE
authors.lastname: Verbeek
authors.forename: J W P M
authors.initials: JW
authors.lastname: van Baal
mesh_terms: D001471:Barrett Esophagus / Q000235:genetics / Q000473:pathology; D001647:Bile Acids and Salts / Q000494:pharmacology*; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D004942:Esophagitis, Peptic / Q000235:genetics; D004947:Esophagus / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015850:Interleukin-6 / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D055262:Mucin-2 / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D055758:SOX9 Transcription Factor / Q000378:metabolism; D013450:Sulfones / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D001647:Bile Acids and Salts; D015850:Interleukin-6; C542359:MIRN143 microRNA, human; C541010:MIRN145 microRNA, human; C546783:MIRN192 microRNA, human; C554846:MIRN194 microRNA, human; C524625:MUC2 protein, human; D035683:MicroRNAs; D055262:Mucin-2; D016328:NF-kappa B; D009570:Nitriles; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D013450:Sulfones; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human
keywords: Barrett's esophagus; bile salt; in vitro model; microRNA; reflux esophagitis