MGMT-B Gene Promoter Hypermethylation in Patients with Inflammatory Bowel Disease - A Novel Finding

  • Mokarram, Pooneh (Department of Biochemistry, Gastroenterohepatology Research Center, Nemazee Hospital, Colorectal Research Center) ;
  • Kavousipour, Soudabeh (Department of Biochemistry, Gastroenterohepatology Research Center, Nemazee Hospital, Colorectal Research Center) ;
  • Sarabi, Mostafa Moradi (Department of Biochemistry, Gastroenterohepatology Research Center, Nemazee Hospital, Colorectal Research Center) ;
  • Mehrabani, Golnosh (Department of Biochemistry, Gastroenterohepatology Research Center, Nemazee Hospital, Colorectal Research Center) ;
  • Fahmidehkar, Mohammad Ali (Department of Biochemistry, Gastroenterohepatology Research Center, Nemazee Hospital, Colorectal Research Center) ;
  • Shamsdin, Seyedeh Azra (Department of Biochemistry, Gastroenterohepatology Research Center, Nemazee Hospital, Colorectal Research Center) ;
  • Alipour, Abbas (Social Medicine Department, Medical Faculty, Mazandaran University of Medical Sciences) ;
  • Naini, Mahvash Alizade (Department of Internal Medicine, School of Medicine, Shiraz University of Medical Sciences)
  • Published : 2015.03.18


Inflammatory bowel disease (IBD) is a disease strongly associated with colorectal cancer (CRC) as a well-known precancerous condition. Alterations in DNA methylation and mutation in K-ras are believed to play an early etiopathogenic role in CRC and may also an initiating event through deregulation of molecular signaling. Epigenetic silencing of APC and SFRP2 in the WNT signaling pathway may also be involved in IBD-CRC. The role of aberrant DNA methylation in precancerous state of colorectal cancer (CRC) is under intensive investigation worldwide. The aim of this study was to investigate the status of promoter methylation of MGMT-B, APC1A and SFRP2 genes, in inflamed and normal colon tissues of patients with IBD compared with control normal tissues. A total of 52 IBD tissues as well as corresponding normal tissues and 30 samples from healthy participants were obtained. We determined promoter methylation status of MGMT-B, SFRP2 and APC1A genes by chemical treatment with sodium bisulfite and subsequent MSP. The most frequently methylated locus was MGMT-B (71%; 34 of 48), followed by SFRP2 (66.6 %; 32 of 48), and APC1A (43.7%; 21 of 48). Our study demonstrated for the first time that hypermethylation of the MGMT-B and the SFRP2 gene promoter regions might be involved in IBD development. Methylation of MGMT-B and SFRP2 in IBD patients may provide a method for early detection of IBD-associated neoplasia.


Inflammatory bowel disease;colorectal cancer;gene methylation


Supported by : Shiraz University of Medical Sciences


  1. Al-Shabanah OA, Hafez MM, Hassan ZK, et al (2014). Methylation of SFRPs and APC genes in ovarian cancer infected with high risk human papillomavirus. Asian Pac J Cancer Prev, 15, 2719-25.
  2. Anastas JN, Moon RT (2013). WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer, 13, 11-26.
  3. Aust DE, Haase M, Dobryden L, et al (2005). Mutations of the BRAF gene in ulcerative colitis-related colorectal carcinoma. Int J Cancer, 115, 673-7.
  4. Azarschab P, Porschen R, Gregor M, Blin N, Holzmann K (2002). Epigenetic control of the E-cadherin gene (CDH1) by CpG methylation in colectomy samples of patients with ulcerative colitis. Genes Chromosomes Cancer, 35, 121-6.
  5. Barker N, Clevers H (2006). Mining the Wnt pathway for cancer therapeutics. Nat Rev Drug Discov, 5, 997-1014.
  6. Baylin SB, Ohm JE (2006). Epigenetic gene silencing in cancer-a mechanism for early oncogenic pathway addiction? Nat Rev Cancer, 6, 107-16.
  7. Bosch LJ, Mongera S, Terhaar Sive Droste JS, et al (2012). Analytical sensitivity and stability of DNA methylation testing in stool samples for colorectal cancer detection. Cell Oncol (Dordr), 35, 309-15.
  8. Brentnall TA, Crispin DA, Rabinovitch PS, et al (1994). Mutations in the p53 gene: an early marker of neoplastic progression in ulcerative colitis. Gastroenterology, 107, 369-78.
  9. Bronner MP, Culin C, Reed JC, Furth EE (1995). The bcl-2 proto-oncogene and the gastrointestinal epithelial tumor progression model. Am J Pathol, 146, 20-6.
  10. Burmer GC, Rabinovitch PS, Haggitt RC, et al (1992). Neoplastic progression in ulcerative colitis: histology, DNA content, and loss of a p53 allele. Gastroenterology, 103, 1602-10.
  11. Chambers WM, Warren BF, Jewell DP, Mortensen NJ (2005). Cancer surveillance in ulcerative colitis. Br J Surg, 92, 928-36.
  12. Cooke J, Zhang H, Greger L, et al (2012). Mucosal genomewide methylation changes in inflammatory bowel disease. Inflamm Bowel Dis, 18, 2128-37.
  13. deVos T, Tetzner R, Model F, et al (2009). Circulating methylated SEPT9 DNA in plasma is a biomarker for colorectal cancer. Clin Chem, 55, 1337-46.
  14. Dhir M, Montgomery EA, Glockner SC, et al (2008). Epigenetic regulation of WNT signaling pathway genes in inflammatory bowel disease (IBD) associated neoplasia. J Gastrointest Surg, 12, 1745-53.
  15. Fang WJ, Zheng Y, Wu LM, et al (2012). Genome-wide analysis of aberrant DNA methylation for identification of potential biomarkers in colorectal cancer patients. Asian Pac J Cancer Prev, 13, 1917-21.
  16. Fleisher AS, Esteller M, Harpaz N, et al (2000). Microsatellite instability in inflammatory bowel disease-associated neoplastic lesions is associated with hypermethylation and diminished expression of the DNA mismatch repair gene, hMLH1. Cancer Res, 60, 4864-8.
  17. Goel GA, Kandiel A, Achkar JP, Lashner B (2011). Molecular pathways underlying IBD-associated colorectal neoplasia: therapeutic implications. Am J Gastroenterol, 106, 719-30.
  18. Gonzalo V, Lozano JJ, Munoz J, et al (2010). Aberrant gene promoter methylation associated with sporadic multiple colorectal cancer. PLoS One, 5, 8777.
  19. Hanauer SB (2006). Inflammatory bowel disease: epidemiology, pathogenesis, and therapeutic opportunities. Inflamm Bowel Dis, 1, 3-9.
  20. Harpaz N, Ward SC, Mescoli C, Itzkowitz SH, Polydorides AD (2013). Precancerous lesions in inflammatory bowel disease. Best Pract Res Clin Gastroenterol, 27, 257-67.
  21. Hartnett L, Egan LJ (2012). Inflammation, DNA methylation and colitis-associated cancer. Carcinogenesis, 33, 723-31.
  22. Hegde ML, Hazra TK, Mitra S (2008). Early steps in the DNA base excision/single-strand interruption repair pathway in mammalian cells. Cell Res, 18, 27-47.
  23. Hegde ML, Hazra TK, Mitra S (2008). Early steps in the DNA base excision/single-strand interruption repair pathway in mammalian cells. Cell Res, 18, 27-47.
  24. Herrinton LJ, Liu L, Levin TR, et al (2012). Incidence and mortality of colorectal adenocarcinoma in persons with inflammatory bowel disease from 1998 to 2010. Gastroenterology, 143, 382-9.
  25. Hong L, Ahuja N (2013). DNA methylation biomarkers of stool and blood for early detection of colon cancer. Genet Test Mol Biomarkers, 17, 401-6.
  26. Issa JP, Ahuja N, Toyota M, Bronner MP, Brentnall TA (2001). Accelerated age-related CpG island methylation in ulcerative colitis. Cancer Res, 61, 3573-7.
  27. Jain SK, Peppercorn MA (1997). Inflammatory bowel disease and colon cancer: a review. Dig Dis, 15, 243-52.
  28. Jawad N, Direkze N, Leedham SJ (2011). Inflammatory bowel disease and colon cancer. Recent Results Cancer Res, 185, 99-115.
  29. Jenke AC, Zilbauer M (2012). Epigenetics in inflammatory bowel disease. Curr Opin Gastroenterol, 28, 577-84.
  30. Kellermayer R (2012). Epigenetics and the developmental origins of inflammatory bowel diseases. Can J Gastroenterol, 26, 909-15.
  31. Konishi K, Shen L, Wang S, et al (2007). Rare CpG island methylator phenotype in ulcerative colitis-associated neoplasias. Gastroenterology, 132, 1254-60.
  32. Lange CP, Campan M, Hinoue T, et al (2012). Genome-scale discovery of DNA-methylation biomarkers for blood-based detection of colorectal cancer. PLoS One, 7, 50266.
  33. Lao VV, Grady WM (2011). Epigenetics and colorectal cancer. Nat Rev Gastroenterol Hepatol, 8, 686-700.
  34. Lin Z, Hegarty JP, Cappel JA, et al (2011). Identification of disease-associated DNA methylation in intestinal tissues from patients with inflammatory bowel disease. Clin Genet, 80, 59-67.
  35. Low D, Mizoguchi A, Mizoguchi E (2013). DNA methylation in inflammatory bowel disease and beyond. World J Gastroenterol, 19, 5238-49.
  36. Matsumura S, Oue N, Ito R, et al (2003). The promoter methylation status of the DNA repair gene O6-methylguanine-DNA methyltransferase in ulcerative colitis. Virchows Arch, 443, 518-23.
  37. Mattar MC, Lough D, Pishvaian MJ, Charabaty A (2011). Current management of inflammatory bowel disease and colorectal cancer. Gastrointest Cancer Res, 4, 53-61.
  38. Mikami T, Yoshida T, Numata Y, et al (2007). Low frequency of promoter methylation of O6-methylguanine DNA methyltransferase and hMLH1 in ulcerative colitisassociated tumors: comparison with sporadic colonic tumors. Am J Clin Pathol, 127, 366-73.
  39. Mokarram P, Naghibalhossaini F, Saberi Firoozi M, et al (2008). Methylenetetrahydrofolate reductase C677T genotype affects promoter methylation of tumor-specific genes in sporadic colorectal cancer through an interaction with folate/ vitamin B12 status. World J Gastroenterol, 14, 3662-71.
  40. Mokarram P, Zamani M, Kavousipour S, et al (2013). Different patterns of DNA methylation of the two distinct O6- methylguanine-DNA methyltransferase (O6-MGMT) promoter regions in colorectal cancer. Mol Biol Rep, 40, 3851-7.
  41. Munkholm P (2003). Review article: the incidence and prevalence of colorectal cancer in inflammatory bowel disease. Aliment Pharmacol Ther, 2, 1-5.
  42. Nakagawa H, Koyama K, Murata Y, et al (2000). APCL, a central nervous system-specific homologue of adenomatous polyposis coli tumor suppressor, binds to p53-binding protein 2 and translocates it to the perinucleus. Cancer Res, 60, 101-5.
  43. Noah TK, Lo YH, Price A, et al (2013). SPDEF functions as a colorectal tumor suppressor by inhibiting beta-catenin activity. Gastroenterology, 144, 1012-23.
  44. Okochi O, Hibi K, Sakai M, et al (2003). Methylation-mediated silencing of SOCS-1 gene in hepatocellular carcinoma derived from cirrhosis. Clin Cancer Res, 9, 5295-8.
  45. Olaru AV, Cheng Y, Agarwal R, et al (2012). Unique patterns of CpG island methylation in inflammatory bowel diseaseassociated colorectal cancers. Inflamm Bowel Dis, 18, 641-8.
  46. Pandurangan AK, Esa NM (2014). Signal transducer and activator of transcription 3-a promising target in colitisassociated cancer. Asian Pac J Cancer Prev, 15, 551-60.
  47. Petronis A, Petroniene R (2000). Epigenetics of inflammatory bowel disease. Gut, 47, 302-6.
  48. Philipp AB, Stieber P, Nagel D, et al (2012). Prognostic role of methylated free circulating DNA in colorectal cancer. Int J Cancer, 131, 2308-19.
  49. Piperi C, Themistocleous MS, Papavassiliou GA, et al (2010). High incidence of MGMT and RARbeta promoter methylation in primary glioblastomas: association with histopathological characteristics, inflammatory mediators and clinical outcome. Mol Med, 16, 1-9.
  50. Psofaki V, Kalogera C, Tzambouras N, et al (2010). Promoter methylation status of hMLH1, MGMT, and CDKN2A/p16 in colorectal adenomas. World J Gastroenterol, 16, 3553-60.
  51. Sangplod P, Kanngurn S, Boonpipattanapong T, Ruangrat P, Sangkhathat S (2014). Expression of BMP6 is associated with its methylation status in colorectal cancer tissue but lacks prognostic significance. Asian Pac J Cancer Prev, 15, 7091-5.
  52. Sato F, Shibata D, Harpaz N, et al (2002). Aberrant methylation of the HPP1 gene in ulcerative colitis-associated colorectal carcinoma. Cancer Res, 62, 6820-2.
  53. Shamsara J, Sharif S, Afsharnezhad S, et al (2009). Association between MGMT promoter hypermethylation and p53 mutation in glioblastoma. Cancer Invest, 27, 825-9.
  54. Summers T, Langan RC, Nissan A, et al (2013). Serum-based DNA methylation biomarkers in colorectal cancer: potential for screening and early detection. J Cancer, 4, 210-6.
  55. Svrcek M, Buhard O, Colas C, et al (2010). Methylation tolerance due to an O6-methylguanine DNA methyltransferase (MGMT) field defect in the colonic mucosa: an initiating step in the development of mismatch repair-deficient colorectal cancers. Gut, 59, 1516-26.
  56. Switzeny OJ, Mullner E, Wagner KH, et al (2012). Vitamin and antioxidant rich diet increases MLH1 promoter DNA methylation in DMT2 subjects. Clin Epigenetics, 4, 19.
  57. Tahara T, Shibata T, Nakamura M, et al (2009). Effect of MDR1 gene promoter methylation in patients with ulcerative colitis. Int J Mol Med, 23, 521-7.
  58. Ullman TA, Itzkowitz SH (2011). Intestinal inflammation and cancer. Gastroenterology, 140, 1807-16.
  59. Wani M, Afroze D, Makhdoomi M, et al (2012). Promoter methylation status of DNA repair gene (hMLH1) in gastric carcinoma patients of the Kashmir valley. Asian Pac J Cancer Prev, 13, 4177-81.
  60. Waris G, Ahsan H (2006). Reactive oxygen species: role in the development of cancer and various chronic conditions. J Carcinog, 5, 14.
  61. Whitehall VL, Walsh MD, Young J, Leggett BA, Jass JR (2001). Methylation of O-6-methylguanine DNA methyltransferase characterizes a subset of colorectal cancer with low-level DNA microsatellite instability. Cancer Res, 61, 827-30.
  62. Widschwendter M, Menon U (2006). Circulating methylated DNA: a new generation of tumor markers. Clin Cancer Res, 12, 7205-8.
  63. Wu JY, Wang J, Lai JC, et al (2008). Association of O6- methylguanine-DNA methyltransferase (MGMT) promoter methylation with p53 mutation occurrence in non-small cell lung cancer with different histology, gender, and smoking status. Ann Surg Oncol, 15, 3272-7.
  64. Yin J, Harpaz N, Tong Y, et al (1993). p53 point mutations in dysplastic and cancerous ulcerative colitis lesions. Gastroenterology, 104, 1633-9.
  65. You J, Nguyen AV, Albers CG, Lin F, Holcombe RF (2008). Wnt pathway-related gene expression in inflammatory bowel disease. Dig Dis Sci, 53, 1013-9.
  66. Zhai R, Zhao Y, Su L, et al (2012). Genome-wide DNA methylation profiling of cell-free serum DNA in esophageal adenocarcinoma and Barrett esophagus. Neoplasia, 14, 29-33.
  67. Zhang W, Bauer M, Croner RS, et al (2007). DNA stool test for colorectal cancer: hypermethylation of the secreted frizzledrelated protein-1 gene. Dis Colon Rectum, 50, 1618-26.
  68. Zisman TL, Bronner MP, Rulyak S, et al (2012). Prospective study of the progression of low-grade dysplasia in ulcerative colitis using current cancer surveillance guidelines. Inflamm Bowel Dis, 18, 2240-6.
  69. Zisman TL, Rubin DT (2008). Colorectal cancer and dysplasia in inflammatory bowel disease. World J Gastroenterol, 14, 2662-9.

Cited by

  1. Sensitive and Noninvasive Detection of Aberrant SFRP2 and MGMT-B Methylation in Iranian Patients with Colon Polyps vol.17, pp.4, 2016,
  2. Epigenetic biomarkers in colorectal cancer: premises and prospects pp.1366-5804, 2016,