Abstract
The human genes MUC2, MUC5AC, MUC5B and MUC6 are clustered on chromosome 11 and encode large secreted gel-forming mucins. The frequent occurrence of their silencing in cancers and the GC-rich structure of their promoters led us to study the influence of epigenetics on their expression. Pre- and post-confluent cells were treated with demethylating agent 5-aza-2′-deoxycytidine and histone deacetylase (HDAC) inhibitor, trichostatin A. Mapping of methylated cytosines was performed by bisulfite-treated genomic DNA sequencing. Histone modification status at the promoters was assessed by chromatin immunoprecipitation assays. Our results indicate that MUC2 was regulated by site-specific DNA methylation associated with establishment of a repressive histone code, whereas hypermethylation of MUC5B promoter was the major mechanism responsible for its silencing. DNA methyltransferase 1 was identified by small interfering RNA approach as a regulator of MUC2 and MUC5B endogenous expression that was potentiated by HDAC2. MUC2 and MUC5B epigenetic regulation was cell-specific, depended on cell differentiation status and inhibited their activation by Sp1. The expression of MUC5AC was rarely influenced by epigenetic mechanisms and methylation of MUC6 promoter was not correlated to its silencing. In conclusion, this study demonstrates the important role for methylation and/or histone modifications in regulating the 11p15 mucin genes in epithelial cancer cells.
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References
Bernacki SH, Nelson AL, Abdullah L, Sheehan JK, Harris A, Davis CW et al. (1999). Mucin gene expression during differentiation of human airway epithelia in vitro. MUC4 and MUC5B are strongly induced. Am J Respir Cell Mol Biol 20: 595–604.
Blache P, Van de Wetering M, Duluc I, Domon C, Berta P, Freund JN et al. (2004). SOX9 is an intestine crypt transcription factor, is regulated by the Wnt pathway, and represses the CDX2 and MUC2 genes. J Cell Biol 166: 37–47.
Copin MC, Buisine MP, Devisme L, Leroy X, Escande F, Gosselin B et al. (2001). Normal respiratory mucosa, precursor lesions and lung carcinomas: differential expression of human mucin genes. Front Biosci 6: D1264–D1275.
De Bustros A, Nelkin BD, Silverman A, Ehrlich G, Poiesz B, Baylin SB . (1988). The short arm of chromosome 11 is a ‘hot spot’ for hypermethylation in human neoplasia. Proc Natl Acad Sci USA 85: 5693–5697.
De Ruijter AJ, Van Gennip AH, Caron HN, Kemp S, Van Kuilenburg AB . (2003). Histone deacetylases (HDACs): characterization of the classical HDAC family. Biochem J 370: 737–749.
Desseyn JL, Aubert JP, Porchet N, Laine A . (2000). Evolution of the large secreted gel-forming mucins. Mol Biol Evol 17: 1175–1184.
Fahrner JA, Eguchi S, Herman JG, Baylin SB . (2002). Dependence of histone modifications and gene expression on DNA hypermethylation in cancer. Cancer Res 62: 7213–7218.
Ferguson M, Henry PA, Currie RA . (2003). Histone deacetylase inhibition is associated with transcriptional repression of the Hmga2 gene. Nucleic Acids Res 31: 3123–3133.
Ghoshal K, Majumder S, Li Z, Dong X, Jacob ST . (2000). Suppression of metallothionein gene expression in a rat hepatoma because of promoter-specific DNA methylation. J Biol Chem 275: 539–547.
Gratchev A, Bohm C, Riede E, Foss HD, Hummel M, Mann B et al. (1998). Regulation of mucin MUC2 gene expression during colon carcinogenesis. Ann NY Acad Sci 859: 180–183.
Gratchev A, Siedow A, Bumke-Vogt C, Hummel M, Foss HD, Hanski ML et al. (2001). Regulation of the intestinal mucin MUC2 gene expression in vivo: evidence for the role of promoter methylation. Cancer Lett 168: 71–80.
Hamada T, Goto M, Tsutsumida H, Nomoto M, Higashi M, Sugai T et al. (2005). Mapping of the methylation pattern of the MUC2 promoter in pancreatic cancer cell lines, using bisulfite genomic sequencing. Cancer Lett 227: 175–184.
Hanski C, Riede E, Gratchev A, Foss HD, Bohm C, Klussmann E et al. (1997). MUC2 gene suppression in human colorectal carcinomas and their metastases: in vitro evidence of the modulatory role of DNA methylation. Lab Invest 77: 685–695.
Herman JG, Baylin SB . (2003). Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med 349: 2042–2054.
Ho JJ, Han SW, Pan PL, Deng G, Kuan SF, Kim YS . (2003). Methylation status of promoters and expression of MUC2 and MUC5AC mucins in pancreatic cancer cells. Int J Oncol 22: 273–279.
Hollingsworth MA, Swanson BJ . (2004). Mucins in cancer: protection and control of the cell surface. Nat Rev Cancer 4: 45–60.
Koo JS, Yoon JH, Gray T, Norford D, Jetten AM, Nettesheim P . (1999). Restoration of the mucous phenotype by retinoic acid in retinoid-deficient human bronchial cell cultures: changes in mucin gene expression. Am J Respir Cell Mol Biol 20: 43–52.
Laird PW . (2003). The power and the promise of DNA methylation markers. Nat Rev 3: 253–266.
Leteurtre E, Gouyer V, Rousseau K, Moreau O, Barbat A, Swallow D et al. (2004). Differential mucin expression in colon carcinoma HT-29 clones with variable resistance to 5-fluorouracil and methotrexate. Biol Cell 96: 145–151.
Mariette C, Perrais M, Leteurtre E, Jonckheere N, Hemon B, Pigny P et al. (2004). 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 377: 701–708.
Mesquita P, Peixoto AJ, Seruca R, Hanski C, Almeida R, Silva F et al. (2003). Role of site-specific promoter hypomethylation in aberrant MUC2 mucin expression in mucinous gastric carcinomas. Cancer Lett 189: 129–136.
Momparler RL . (2003). Cancer epigenetics. Oncogene 22: 6479–6483.
Perrais M, Pigny P, Buisine MP, Porchet N, Aubert JP, Van Seuningen-Lempire I . (2001a). Aberrant expression of human mucin gene MUC5B in gastric carcinoma and cancer cells. Identification and regulation of a distal promoter. J Biol Chem 276: 15386–15396.
Perrais M, Pigny P, Copin MC, Aubert JP, Van Seuningen I . (2002). 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 277: 32258–32267.
Perrais M, Pigny P, Ducourouble MP, Petitprez D, Porchet N, Aubert JP et al. (2001b). Characterization of human mucin gene MUC4 promoter: importance of growth factors and proinflammatory cytokines for its regulation in pancreatic cancer cells. J Biol Chem 276: 30923–30933.
Piessen G, Jonckheere N, Vincent A, Hemon B, Ducourouble MP, Copin MC et al. (2007). 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 402: 81–91.
Pigny P, Guyonnet-Duperat V, Hill AS, Pratt WS, Galiegue-Zouitina S, D'Hooge MC et al. (1996). Human mucin genes assigned to 11p15.5: identification and organization of a cluster of genes. Genomics 38: 340–352.
Reis CA, Sorensen T, Mandel U, David L, Mirgorodskaya E, Roepstorff P et al. (1998). Development and characterization of an antibody directed to an alpha-N-acetyl-D-galactosamine glycosylated MUC2 peptide. Glycoconj J 15: 51–62.
Rousseau K, Wickstrom C, Whitehouse DB, Carlstedt I, Swallow DM . (2003). New monoclonal antibodies to non-glycosylated domains of the secreted mucins MUC5B and MUC7. Hybrid Hybridomics 22: 293–299.
Siedow A, Szyf M, Gratchev A, Kobalz U, Hanski ML, Bumke-Vogt C et al. (2002). De novo expression of the Muc2 gene in pancreas carcinoma cells is triggered by promoter demethylation. Tumour Biol 23: 54–60.
Sylvester PA, Myerscough N, Warren BF, Carlstedt I, Corfield AP, Durdey P et al. (2001). Differential expression of the chromosome 11 mucin genes in colorectal cancer. J Pathol 195: 327–335.
Van Seuningen I, Perrais M, Pigny P, Porchet N, Aubert JP . (2000). Sequence of the 5′-flanking region and promoter activity of the human mucin gene MUC5B in different phenotypes of colon cancer cells. Biochem J 348: 675–686.
Van Seuningen I, Pigny P, Perrais M, Porchet N, Aubert JP . (2001). Transcriptional regulation of the 11p15 mucin genes. Towards new biological tools in human therapy, in inflammatory diseases and cancer? Front Biosci 6: D1216–D1234.
Yamada N, Hamada T, Goto M, Tsutsumida H, Higashi M, Nomoto M et al. (2006). MUC2 expression is regulated by histone H3 modification and DNA methylation in pancreatic cancer. Int J Cancer 119: 1850–1857.
Zhu P, Huber E, Kiefer F, Gottlicher M . (2004). Specific and redundant functions of histone deacetylases in regulation of cell cycle and apoptosis. Cell Cycle 3: 1240–1242.
Acknowledgements
We are indebted to Michel Crépin (Laboratoire de Biochimie-Biologie Moléculaire, CHRU-Eurasanté, Lille, France), Dominique Demeyer and Marie-Paule Ducourouble for their excellent technical help. We thank Dr T Lesuffleur for the kind gift of colonic HT-29 5F7 cells and Dr CA Reis (IPATIMUP, Porto, Portugal) for the gift of anti-MUC2 (PMH1) and anti-MUC6 (CLH5) antibodies. Audrey Vincent is the recipient of a Conseil Régional Nord-Pas de Calais and the Institut National de la Santé et de la Recherche Médicale PhD fellowship. This work was supported by a grant from l'Association pour la Recherche sur le Cancer (Isabelle Van Seuningen, Grant no. 3872).
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Dedicated to Dr Jean-Pierre Aubert (DR1 INSERM) who passed away on 3 September, 2005. It is an immense loss for us, as the director of our laboratory, and for the mucin field scientific community, who appreciated him for his human qualities and as the discoverer of three mucin genes in the 1990s.
Sequences submitted to Genbank: AY500284, AY370683, AY458429.
Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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Vincent, A., Perrais, M., Desseyn, JL. et al. Epigenetic regulation (DNA methylation, histone modifications) of the 11p15 mucin genes (MUC2, MUC5AC, MUC5B, MUC6) in epithelial cancer cells. Oncogene 26, 6566–6576 (2007). https://doi.org/10.1038/sj.onc.1210479
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DOI: https://doi.org/10.1038/sj.onc.1210479
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