Original communication| Volume 143, ISSUE 1, P58-71, January 2008

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Activation of MUC1 mucin expression by bile acids in human esophageal adenocarcinomatous cells and tissues is mediated by the phosphatidylinositol 3-kinase


      In esophageal adenocarcinoma, MUC1 mucin expression increases in early stages of the carcinogenetic sequence, during which bile reflux has been identified as a major carcinogen. However, no link between MUC1 overexpression and the presence of bile acids in the reflux has been established so far, and molecular mechanisms regulating MUC1 expression during esophageal carcinogenetic sequence are unknown. Our aim was to identify (1) the bile acids able to upregulate MUC1 expression in esophageal cancer cells and mucosal samples, (2) the regulatory regions in MUC1 promoter responsive to bile acids, and (3) the signaling pathway(s) involved in this regulation.


      MUC1 mRNA and mucin expression were studied by the means of real-time reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry, both in the human esophageal OE33 adenocarcinoma cell line and in an ex vivo explant model. MUC1 promoter was cloned and transcription regulation was studied by transient cell transfection to identify the bile acid–responsive regions. Signaling pathways involved were identified using specific pharmacologic inhibitors and siRNA approach.


      Taurocholic, taurodeoxycholic, taurochenodeoxycholic, glycocholic, sodium glycocholate, and deoxycholic bile acids upregulated MUC1 mRNA and protein expression. The highest induction was obtained with deoxycholic and taurocholic acids in both cellular and explant models. The bile acid–mediated upregulation of MUC1 transcription occurs at the promoter level, with responsive elements located in the -1472/-234 region of the promoter, and involves the phosphatidylinositol 3-kinase signaling pathway.


      Bile acids induce MUC1 mucin overexpression in human esophageal adenocarcinoma cells and tissues by activating its transcription through a process involving phosphatidylinositol 3-kinase.
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        • DeMeester S.R.
        Adenocarcinoma of the esophagus and cardia: a review of the disease and its treatment.
        Ann Surg Oncol. 2006; 13: 2-30
        • Mariette C.
        • Finzi L.
        • Piessen G.
        • Van Seuningen I.
        • Triboulet J.-P.
        Esophageal carcinoma: prognostic differences between squamous cell carcinoma and adenocarcinoma.
        World J Surg. 2005; 29: 39-45
        • Wild C.P.
        • Hardie L.J.
        Reflux, Barrett’s oesophagus and adenocarcinoma: burning questions.
        Nature Rev Cancer. 2003; 3: 676-685
        • Ye W.
        • Chow W.H.
        • Lagergren J.
        • Yin L.
        • Nyren O.
        Risk of adenocarcinomas of the esophagus and gastric cardia in patients with gastroesophageal reflux diseases and after antireflux surgery.
        Gastroenterology. 2001; 121: 1286-1293
        • Warson C.
        • Van De Bovenkamp J.H.
        • Korteland-Van Male A.M.
        • Buller H.A.
        • Einerhand A.W.
        • Ectors N.L.
        • et al.
        Barrett’s esophagus is characterized by expression of gastric-type mucins (MUC5AC, MUC6) and TFF peptides (TFF1 and TFF2), but the risk of carcinoma development may be indicated by the intestinal-type mucin, MUC2.
        Hum Pathol. 2002; 33: 660-668
        • Gillen P.
        • Thornton J.
        • Byrne P.J.
        • Walsh T.N.
        • Hennessy T.P.
        Implications of upright gastro-oesophageal reflux.
        Br J Surg. 1994; 81: 239-240
        • Jankowski J.A.
        • Harrison R.F.
        • Perry I.
        • Balkwill F.
        • Tselepis C.
        Barrett’s metaplasia.
        Lancet. 2000; 356: 2079-2085
        • Sato T.
        • Miwa K.
        • Sahara H.
        • Segawa M.
        • Hattori T.
        The sequential model of Barrett’s esophagus and adenocarcinoma induced by duodeno-esophageal reflux without exogenous carcinogens.
        Anticancer Res. 2002; 22: 39-44
        • Goldstein S.R.
        • Yang G.Y.
        • Curtis S.K.
        • Reuhl K.R.
        • Liu B.C.
        • Mirvish S.
        • et al.
        Development of esophageal metaplasia and adenocarcinoma in a rat surgical model without the use of a carcinogen.
        Carcinogenesis. 1997; 18: 2265-2270
        • Montgomery E.
        • Goldblum J.R.
        • Greenson J.K.
        • Haber M.M.
        • Lamps L.W.
        • Lauwers G.Y.
        • et al.
        Dysplasia as a predictive marker for invasive carcinoma in Barrett esophagus: a follow-up study based on 138 cases from a diagnostic variability study.
        Hum Pathol. 2001; 32: 379-388
        • Hollingsworth M.A.
        • Swanson B.J.
        Mucins in cancer: protection and control of the cell surface.
        Nat Rev Cancer. 2004; 4: 45-60
        • Arul G.S.
        • Moorghen M.
        • Myerscough N.
        • Alderson D.A.
        • Spicer R.D.
        • Corfield A.P.
        Mucin gene expression in Barrett’s oesophagus: an in situ hybridisation and immunohistochemical study.
        Gut. 2000; 47: 753-761
        • Corfield A.P.
        • Myerscough N.
        • Longman R.
        • Sylvester P.
        • Arul S.
        • Pignatelli M.
        Mucins and mucosal protection in the gastrointestinal tract: new prospects for mucins in the pathology of gastrointestinal disease.
        Gut. 2000; 47: 589-594
        • Gendler S.J.
        MUC1: the Renaissance molecule.
        J Mammary Gland Biol Neoplasia. 2001; 6: 339-353
        • von Mensdorff-Pouilly S.
        • Snijdewint F.G.M.
        • Verstraeten A.A.
        • Verheijen R.H.M.
        • Kenemans P.
        Human MUC1 mucin: a multifaceted glycoprotein.
        Int J Biol Markers. 2000; 15: 343-356
        • Baldus S.E.
        • Engelmann K.
        • Hanisch F.-G.
        MUC1 and the MUCs: a family of human mucins with impact in cancer biology.
        Crit Rev Clin Lab Sci. 2004; 41: 189-231
        • Dube D.H.
        • Prescher J.A.
        • Quang C.N.
        • Bertozzi C.R.
        Probing mucin-type O-linked glycosylation in living animals.
        Proc Natl Acad Sci U S A. 2006; 103: 4819-4824
        • Guillem P.
        • Billeret V.
        • Buisine M.P.
        • Flejou J.-F.
        • Lecomte-Houcke M.
        • Degand P.
        • et al.
        Mucin gene expression and cell differentiation in human normal, premalignant and malignant esophagus.
        Int J Cancer. 2000; 88: 856-861
        • Glickman J.N.
        • Shahsafaei A.
        • Odze R.D.
        Mucin core peptide expression can help differentiate Barrett’s oesophagus from intestinal metaplasia of the stomach.
        Am J Surg Pathol. 2003; 27: 1357-1365
        • Bax D.A.
        • Einerhand A.W.C.
        • van Dekken H.
        • Blok P.
        • Siersema P.D.
        • Kuipers E.J.
        • et al.
        MUC4 is increased in high grade intraepithelial neoplasia in Barrett’s oesophagus and is associated with a proapoptotic Bax to Bcl-2 ratio.
        J Clin Pathol. 2004; 57: 1267-1272
        • Mariette C.
        • Perrais M.
        • Leteurtre E.
        • Jonckheere N.
        • Hemon B.
        • Pigny P.
        • et al.
        Transcriptional regulation of human mucin MUC4 by bile acids in oesophageal cancer cells is promoter-dependent and involves activation of the phosphatidylinositol 3-kinase signalling pathway.
        Biochem J. 2004; 377: 701-708
        • Song S.
        • Byrd J.C.
        • Koo J.S.
        • Bresalier R.S.
        Bile acids induce MUC2 overexpression in human colon carcinoma cells.
        Cancer. 2005; 103: 1606-1614
        • Piessen G.
        • Jonckheere N.
        • Vincent A.
        • Hemon B.
        • Ducourouble M.-P.
        • Copin M.-C.
        • et al.
        Regulation of the human mucin MUC4 by taurodeoxycholic and taurochenodeoxycholic bile acids in oesophageal cancer cells is mediated by hepatocyte nuclear factor 1alpha.
        Biochem J. 2007; 402: 81-91
        • Van Seuningen I.
        • Perrais M.
        • Pigny P.
        • Porchet N.
        • Aubert J.-P.
        Sequence of the 5’-flanking region and promoter activity of the human mucin gene MUC5B in different phenotypes of colon cancer cells.
        Biochem J. 2000; 348: 675-686
        • Lan M.
        • Batra S.K.
        • Qi W.-N.
        • Metzgar R.S.
        • Hollingsworth M.A.
        Cloning and sequencing of a human pancreatic tumor mucin cDNA.
        J Biol Chem. 1990; 265: 15294-15299
        • Mesquita P.
        • Jonckheere N.
        • Almeida R.
        • et al.
        Human MUC2 mucin gene is transcriptionally regulated by Cdx homeodomain proteins in gastrointestinal carcinoma cell lines.
        J Biol Chem. 2003; 278: 51549-51556
        • Matuoka K.
        • Chen K.Y.
        • Takenawa T.
        A positive role of phosphatidylinositol 3-kinase in aging phenotype expression in cultured human diploid fibroblasts.
        Arch Gerontol Geriatr. 2003; 36: 203-219
        • McIlhinney R.A.J.
        • Patel S.
        • Gore M.E.
        Monoclonal antibodies recognizing epitopes carried on both glycolipids and glycoproteins of the human milk fat globule membrane.
        Biochem J. 1985; 227: 155-162
        • Perrais M.
        • Pigny P.
        • Copin M.-C.
        • Aubert J.-P.
        • Van Seuningen I.
        Induction of MUC2 and MUC5AC mucins by factors of the epidermal growth factor (EGF) family is mediated by EGF receptor/Ras/Raf/extracellular signal-regulated kinase cascade and Sp1.
        J Biol Chem. 2002; 277: 32258-32267
        • Beuers U.
        Effects of bile acids on hepatocellular signaling and secretion.
        Yale J Biol Med. 1997; 70: 341-346
        • Brady L.M.
        • Beno D.W.
        • Davis B.H.
        Bile acid stimulation of early growth response gene and mitogen-activated protein kinase is protein kinase C-dependent.
        Biochem J. 1996; 316: 765-769
        • Rust C.
        • Karnitz L.M.
        • Paya C.V.
        • Moscat J.
        • Simari R.D.
        • Gores G.J.
        The bile acid taurochenodeoxycholate activates a phosphatidylinositol 3-kinase-dependent survival signaling cascade.
        J Biol Chem. 2000; 275: 20210-20216
        • Jaiswal K.
        • Tello V.
        • Lopez-Guzman C.
        • Nwariaku F.
        • Anthony T.
        • Sarosi Jr., G.A.
        Bile salt exposure causes phosphatidyl-inositol-3-kinase-mediated proliferation in a Barrett’s adenocarcinoma cell line.
        Surgery. 2004; 136: 160-168
        • Raina D.
        • Kharbanda S.
        • Kufe D.
        The MUC1 oncoprotein activates the anti-apoptotic phosphoinositide 3-kinase/Akt and Bcl-xL pathways in rat 3Y1 fibroblasts.
        J Biol Chem. 2004; 279: 20607-20612
        • Chang F.
        • Lee J.T.
        • Navolanic P.M.
        • Steelman L.S.
        • Shelton J.G.
        • Blalock W.L.
        • et al.
        Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy.
        Leukemia. 2003; 17: 590-603
        • Tullai J.W.
        • Schaffer M.E.
        • Mullenbrock S.
        • Kasif S.
        • Cooper G.M.
        Identification of transcription factor binding sites upstream of human genes regulated by the phosphatidylinositol 3-kinase and MEK/ERK signaling pathways.
        J Biol Chem. 2004; 279: 20167-20177
        • Kim S.
        • Domon-Dell C.
        • Wang Q.
        • Chung D.H.
        • Di Cristofano A.
        • Pandolfi P.P.
        • et al.
        PTEN and TNF-alpha regulation of the intestinal-specific Cdx-2 homeobox gene through a PI3K, PKB/Akt, and NF-kappaB-dependent pathway.
        Gastroenterology. 2002; 123: 1163-1178
        • Abdel-Latif M.M.
        • O’Riordan J.
        • Windle H.J.
        • Carton E.
        • Ravi N.
        • Kelleher D.
        • et al.
        NF-kappaB activation in esophageal adenocarcinoma: relationship to Barrett’s metaplasia, survival, and response to neoadjuvant chemoradiotherapy.
        Ann Surg. 2004; 239: 491-500
        • Jenkins G.J.
        • Harries K.
        • Doak S.H.
        • Wilmes A.
        • Griffiths A.P.
        • Baxter J.N.
        • et al.
        The bile acid deoxycholic acid (DCA) at neutral pH activates NF-kappaB and induces IL-8 expression in oesophageal cells in vitro.
        Carcinogenesis. 2004; 25: 317-323
        • Jung D.
        • Kullak-Ublick G.A.
        Hepatocyte nuclear factor 1 alpha: a key mediator of the effect of bile acids on gene expression.
        Hepatology. 2003; 37: 622-631
        • Kazumori H.
        • Ishihara S.
        • Rumi M.A.
        • Kadowaki Y.
        • Kinoshita Y.
        Bile acids directly augment caudal related homeobox gene Cdx2 expression in oesophageal keratinocytes in Barrett’s epithelium.
        Gut. 2006; 55: 16-25
        • Kaur B.S.
        • Ouatu-Lascar R.
        • Omary M.B.
        • Triadafilopoulos G.
        Bile salts induce or blunt cell proliferation in Barrett’s esophagus in an acid-dependent fashion.
        Am J Physiol. 2000; 278: G1000-G1009
        • Garewal H.
        • Bernstein H.
        • Bernstein C.
        • Sampliner R.
        • Payne C.
        Reduced bile acid-induced apoptosis in “normal” colorectal mucosa: a potential biological marker for cancer risk.
        Cancer Res. 1996; 56: 1480-1483
        • Fein M.
        • Fuchs K.H.
        • Stopper H.
        • Diem S.
        • Herderich M.
        Duodenogastric reflux and foregut carcinogenesis: analysis of duodenal juice in a rodent model of cancer.
        Carcinogenesis. 2000; 21: 2079-2084
        • Gillen P.
        • Keeling P.
        • Byrne P.J.
        • Healy M.
        • O’Moore R.R.
        • Hennessy T.P.
        Implication of duodenogastric reflux in the pathogenesis of Barrett’s oesophagus.
        Br J Surg. 1988; 75: 540-543
        • Nehra D.
        • Howell P.
        • Williams C.P.
        • Pye J.K.
        • Beynon J.
        Toxic bile acids in gastro-oesophageal reflux disease: influence of gastric acidity.
        Gut. 1999; 44: 598-602
        • Holt P.R.
        Competitive inhibition of intestinal bile salt absorption in the rat.
        Am J Physiol. 1966; 210: 63563-63569
        • Cohen B.I.
        • Raicht R.F.
        • Deschner E.E.
        • Takahashi M.
        • Sarwal A.N.
        • Fazzini E.
        Effect of cholic acid feeding on N-methyl-N-nitrosourea-induced colon tumors and cell kinetics in rats.
        J Natl Cancer Inst. 1980; 64: 573-578
        • Martinez J.D.
        • Stratagoules E.D.
        • LaRue J.M.
        • Powell A.A.
        • Gause P.R.
        • Craven M.T.
        • et al.
        Different bile acids exhibit distinct biological effects: the tumor promoter deoxycholic acid induces apoptosis and the chemopreventive agent ursodeoxycholic acid inhibits cell proliferation.
        Nutr Cancer. 1998; 31: 111-118
        • Marchetti M.C.
        • Migliorati G.
        • Moraca R.
        • Riccardi C.
        • Nicoletti I.
        • Fabiani R.
        • et al.
        Possible mechanisms involved in apoptosis of colon tumor cell lines induced by deoxycholic acid, short-chain fatty acids, and their mixtures.
        Nutr Cancer. 1997; 28: 74-80
        • Bernstein C.
        • Bernstein H.
        • Garewal H.
        • Dinning P.
        • Jabi R.
        • Sampliner R.E.
        • et al.
        A bile acid-induced apoptosis assay for colon cancer risk and associated quality control studies.
        Cancer Res. 1999; 59: 2353-2357
        • Sital R.
        • Kusters J.
        • De Rooij F.
        • Kuipers E.
        • Siersema P.
        Bile acids and Barrett’s oesophagus: a sine qua non or coincidence?.
        Scand J Gastroenterol. 2006; 243: 11-17
        • Pera M.
        • Trastek V.F.
        • Carpenter H.A.
        • Fernandez P.L.
        • Cardesa A.
        • Mohr U.
        • Pairolero P.C.
        Influence of pancreatic and biliary reflux on the development of esophageal carcinoma.
        Ann Thorac Surg. 1993; 55: 1386-1392
        • Phillips W.A.
        • Russell S.E.
        • Ciavarella M.L.
        • Choong D.Y.
        • Montgomery K.G.
        • Smith K.
        • et al.
        Mutation analysis of PIK3CA and PIK3CB in esophageal cancer and Barrett’s esophagus.
        Int J Cancer. 2006; 118: 2644-2646
        • Vona-Davis L.
        • Frankenberry K.
        • Cunningham C.
        • Riggs D.R.
        • Jackson B.J.
        • Szwerc M.F.
        • et al.
        MAPK and PI3K inhibition reduces proliferation of Barrett’s adenocarcinoma in vitro.
        J Surg Res. 2005; 127: 53-58
        • Fresno Vara J.A.
        • Casado E.
        • de Castro J.
        • Cejas P.
        • Belda-Iniesta C.
        • Gonzalez-Baron M.
        PI3K/Akt signalling pathway and cancer.
        Cancer Treat Rev. 2004; 30: 193-204