Abstract
Abnormal gastro-oesophageal reflux and bile acids have been linked to the presence of Barrett's oesophageal premalignant lesion associated with an increase in mucin-producing goblet cells and MUC4 mucin gene overexpression. However, the molecular mechanisms underlying the regulation of MUC4 by bile acids are unknown. Since total bile is a complex mixture, we undertook to identify which bile acids are responsible for MUC4 up-regulation by using a wide panel of bile acids and their conjugates. MUC4 apomucin expression was studied by immunohistochemistry both in patient biopsies and OE33 oesophageal cancer cell line. MUC4 mRNA levels and promoter regulation were studied by reverse transcriptase-PCR and transient transfection assays respectively. We show that among the bile acids tested, taurocholic, taurodeoxycholic, taurochenodeoxycholic and glycocholic acids and sodium glycocholate are strong activators of MUC4 expression and that this regulation occurs at the transcriptional level. By using specific pharmacological inhibitors of mitogen-activated protein kinase, phosphatidylinositol 3-kinase, protein kinase A and protein kinase C, we demonstrate that bile acid-mediated up-regulation of MUC4 is promoter-specific and mainly involves activation of phosphatidylinositol 3-kinase. This new mechanism of regulation of MUC4 mucin gene points out an important role for bile acids as key molecules in targeting MUC4 overexpression in early stages of oesophageal carcinogenesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenocarcinoma / enzymology
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Adenocarcinoma / genetics*
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Adenocarcinoma / metabolism
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Adenocarcinoma / pathology
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Bile Acids and Salts / pharmacology
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Bile Acids and Salts / physiology*
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Biomarkers, Tumor / biosynthesis
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Biomarkers, Tumor / genetics
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Cell Line, Tumor
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Enzyme Activation / physiology
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Esophageal Neoplasms / enzymology
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Esophageal Neoplasms / genetics*
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Esophageal Neoplasms / metabolism
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Esophageal Neoplasms / pathology
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Gastric Mucins / genetics
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Gene Expression Regulation / drug effects
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Gene Expression Regulation / physiology
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Humans
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Mucin-1 / biosynthesis
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Mucin-4
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Mucins / biosynthesis
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Mucins / genetics*
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Mucous Membrane / chemistry
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Mucous Membrane / metabolism
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Peptide Fragments / biosynthesis
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Phosphatidylinositol 3-Kinases / metabolism*
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Promoter Regions, Genetic / drug effects
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Promoter Regions, Genetic / genetics
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Promoter Regions, Genetic / physiology*
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RNA, Messenger / biosynthesis
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RNA, Messenger / metabolism
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Signal Transduction / drug effects
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Signal Transduction / physiology*
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Stomach Neoplasms / enzymology
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Stomach Neoplasms / genetics*
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Stomach Neoplasms / metabolism
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Stomach Neoplasms / pathology
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Transcription, Genetic / drug effects
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Transcription, Genetic / physiology*
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Up-Regulation / drug effects
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Up-Regulation / physiology
Substances
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Bile Acids and Salts
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Biomarkers, Tumor
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Gastric Mucins
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MUC1 tandem repeat peptide
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MUC4 protein, human
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Mucin-1
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Mucin-4
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Mucins
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Peptide Fragments
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RNA, Messenger
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apomucin
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Phosphatidylinositol 3-Kinases