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Molecular Biology Reports

, Volume 39, Issue 5, pp 6171–6178 | Cite as

Epigenetic and genetic analysis of WNT signaling pathway in sporadic colorectal cancer patients from Iran

  • Fakhraddin NaghibalhossainiEmail author
  • Mozhdeh Zamani
  • Pooneh Mokarram
  • Islam Khalili
  • Mozhgan Rasti
  • Zohreh Mostafavi-pour
Article

Abstract

The WNT signaling is deregulated in most human colorectal cancers (CRC). Promoter methylation has been proposed as an alternative mechanism to inactivate genes in tumors. To gain insight into the methylation silencing of the WNT pathway during colorectal carcinogenesis, we examined the aberrant methylation profile of four genes, APC, Axin1, Axin2, and GSK3β in an unselected series of 112 sporadic colorectal tumors by methylation specific PCR. It has been suggested that the Axin2 C148T SNP is associated with the risk of developing certain types of cancers. To assess the contribution of Axin2 SNP to CRC susceptibility, we examined the Axin2 C148T genotype in CRC patients and 170 healthy controls by PCR-RFLP. The frequency of CRCs with at least one gene methylated was 18.75%. Promoter methylation of Axin2 and APC genes was detected in 7.1 and 11.9% of tumors, respectively. No aberrant methylation was found in Gsk3β and Axin1 gene in these tumor series. The methylation status of APC had no significant association with clinical parameters. But, promoter methylation of Axin2 was sex-related, occurring more frequently in females (P = 0.002). The frequency of Axin2 C148T genotypes were similar in patients and controls. Moreover, we observed no association between the Axin2 SNP and risk of CRC in patients stratified by age, sex, and smoking status. However, the heterozygote CT genotype was associated with a reduced CRC risk in distal patients compared with proximal patients (OR = 0.3; 95% CI 0.1–0.9, P = 0.04). Our findings indicate that Axin1 and GSK3β methylation play a minor role in colorectal carcinogenesis.

Keywords

Colon cancer WNT signaling AXIN2 P50S SNP Methylation 

Notes

Acknowledgments

This study was part of the dissertation of Mozhdeh Zamani, submitted to Shiraz University of Medical Sciences in partial fulfillment of the requirements for the M.Sc. in biochemistry. This work was supported by a grant from the Vice Chancellor for Research, Shiraz University of Medical Sciences.

References

  1. 1.
    Malekzadeh R, Bishehsari F, Mahdavinia M, Ansari R (2009) Epidemiology and molecular genetics of colorectal cancer in Iran: a review. Arch Iran Med 12:161–169PubMedGoogle Scholar
  2. 2.
    Giles RH, van Es JH, Clevers H (2003) Caught up in a Wnt storm: Wnt signaling in cancer. Biochim Biophys Acta 1653:1–24. doi: 10.1016/S0304-419X(03)00005-2 PubMedGoogle Scholar
  3. 3.
    Thorstensen L, Lind GE, Løvig T, Diep CB, Meling GI, Rognum TO, Lothe RA (2005) Genetic and epigenetic changes of components affecting the WNT pathway in colorectal carcinomas stratified by microsatellite instability. Neoplasia 7:99–108. doi: 10.1593/neo.04448 PubMedCrossRefGoogle Scholar
  4. 4.
    Angers S, Moon RT (2009) Proximal events in Wnt signal transduction. Natl Rev Mol Cell Biol 10:468–477. doi: 10.1038/nrm2717 Google Scholar
  5. 5.
    Kimelman D, Xu W (2006) β-Catenin destruction complex: insights and questions from a structural perspective. Oncogene 25:7482–7491. doi: 10.1038/sj.onc.1210055 PubMedCrossRefGoogle Scholar
  6. 6.
    Behrens J, Lustig B (2004) The Wnt connection to tumorigenesis. Int J Dev Biol 48:477–487. doi: 10.1387/ijdb.041815jb PubMedCrossRefGoogle Scholar
  7. 7.
    Narayan S, Roy D (2003) Role of APC and DNA mismatch repair genes in the development of colorectal cancers. Mol Cancer 2:41. doi: 10.1186/1476-4598-2-41 PubMedCrossRefGoogle Scholar
  8. 8.
    Hiltunen MO, Alhonen L, Koistinaho J, Myohanen S, Paakkonen M, Marin S et al (1997) Hypermethylation of the APC (adenomatous polyposis coli) gene promoter region in human colorectal carcinoma. Int J Cancer 70:644–648. doi: 10.1002/(SICI)1097-0215(19970317) PubMedCrossRefGoogle Scholar
  9. 9.
    Feinberg AP (2004) The epigenetics of cancer etiology. Semin Cancer Biol 14:427–432. doi: 10.1016/j.semcancer.2004.06.005 PubMedCrossRefGoogle Scholar
  10. 10.
    Shimizu Y, Ikeda S, Fujimori M, Kodama S, Nakahara M, Okajima M, Asahara T (2002) Frequent alterations in the Wnt signaling pathway in colorectal cancer with microsatellite instability. Genes Chromosomes Cancer 33:73–81. doi: 10.1002/gcc.1226 PubMedCrossRefGoogle Scholar
  11. 11.
    Liu W, Dong X, Mai M, Seelan RS, Taniguchi K, Krishnadath KK et al (2000) Mutations in AXIN2 cause colorectal cancer with defective mismatch repair by activating beta-catenin/TCF signalling. Nat Genet 26:146–147. doi: 10.1038/79859 PubMedCrossRefGoogle Scholar
  12. 12.
    Kikuchi A (1999) Modulation of Wnt signaling by axin and axil. Cytokine Growth Factor Rev 10:255–265PubMedCrossRefGoogle Scholar
  13. 13.
    Hughes TA, Brady HJM (2005) Cross-talk between pRb/E2F and Wnt/b-catenin pathways: E2F1 induces AXIN2 leading to repression of Wnt signalling and to increased cell death. Exp Cell Res 303:32–46. doi: 10.1016/j.yexcr.2004.09.014 PubMedCrossRefGoogle Scholar
  14. 14.
    Kanzaki H, Ouchida M, Hanafusa H, Yano M, Suzuki H, Aoe M et al (2006) Single nucleotide polymorphism of the AXIN2 gene is preferentially associated with human lung cancer risk in a Japanese population. Int J Mol Med 18:279–284PubMedGoogle Scholar
  15. 15.
    Wang X, Goode EL, Fredericksen ZS, Vierkant RA, Pankratz VS, Liu-Mares W et al (2008) Association of genetic variation in genes implicated in the beta-catenin destruction complex with risk of breast cancer. Cancer Epidemiol Biomarkers Prev 17:2101–2108. doi: 10.1158/1055-9965.EPI-08-0134 PubMedCrossRefGoogle Scholar
  16. 16.
    Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B, Kinzler KW (1997) Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. Science 275:1787–1790. doi: 10.1126/science.275.5307.1787 PubMedCrossRefGoogle Scholar
  17. 17.
    Lin SY, Yeh KT, Chen WT, Chen HC, Chen ST, Chiou HY, Chang JG (2004) Promoter CpG methylation of tumor suppressor genes in colorectal cancer and its relationship to clinical features. Oncol Rep 11:341–348PubMedGoogle Scholar
  18. 18.
    Mokarram P, Naghibalhossaini F, Saberi Firoozi M, Hosseini SV, Izadpanah A, Salahi H 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 14:3662–3671. doi: 10.3748/wjg.14.3662 PubMedCrossRefGoogle Scholar
  19. 19.
    Naghibalhossaini F, Mokarram P, Khalili I, Vasei M, Hosseini SV, Ashktorab H et al (2010) MTHFR C677T and A1298C variant genotypes and the risk of microsatellite instability among Iranian colorectal cancer patients. Cancer Genet Cytogenet 197:142–151. doi: 10.1016/j.cancergencyto.2009.11.014 PubMedCrossRefGoogle Scholar
  20. 20.
    Esteller M, Sparks A, Toyota M, Sanchez-Cespedes M, Capella G, Peinado MA et al (2000) Analysis of adenomatous polyposis coli promoter hypermethylation in human cancer. Cancer Res 60:4366–4371PubMedGoogle Scholar
  21. 21.
    Fu X, Li J, Tian X, Zhang Y (2009) Hypermethylation of APC promoter 1A is associated with moderate activation of Wnt signalling pathway in a subset of colorectal serrated adenomas. Histopathology 55:554–563. doi: 10.1111/j.1365-2559.2009.03411.x PubMedCrossRefGoogle Scholar
  22. 22.
    Kim JC, Choi JS, Roh SA, Cho DH, Kim TW, Kim YS (2010) Promoter methylation of specific genes is associated with the phenotype and progression of colorectal adenocarcinomas. Ann Surg Oncol 17:1767–1776. doi: 10.1245/s10434-009-0901-y PubMedCrossRefGoogle Scholar
  23. 23.
    Lee BB, Lee EJ, Jung EH, Chun HK, Chang DK, Song SY et al (2009) Aberrant methylation of APC, MGMT, RASSF2A, and Wif-1 genes in plasma as a biomarker for early detection of colorectal cancer. Clin Cancer Res 15:6185–6191. doi: 10.1158/1078-0432.CCR-09-0111 PubMedCrossRefGoogle Scholar
  24. 24.
    Chen SP, Chiu SC, Wu CC, Lin SZ, Kang JC, Chen YL et al (2009) The association of methylation in the promoter of APC and MGMT and the prognosis of Taiwanese CRC patients. Genet Test Mol Biomarkers 13:67–71. doi: 10.1089/gtmb.2008.0045 PubMedCrossRefGoogle Scholar
  25. 25.
    Derks S, Postma C, Carvalho B, van den Bosch SM, Moerkerk PT, Herman JG et al (2008) Integrated analysis of chromosomal, microsatellite and epigenetic instability in colorectal cancer identifies specific associations between promoter methylation of pivotal tumour suppressor and DNA repair genes and specific chromosomal alterations. Carcinogenesis 29:434–439. doi: 10.1093/carcin/bgm270 PubMedCrossRefGoogle Scholar
  26. 26.
    Koinuma K, Yamashita Y, Liu W, Hatanaka H, Kurashina K, Wada T et al (2006) Epigenetic silencing of AXIN2 in colorectal carcinoma with microsatellite instability. Oncogene 25:139–146. doi: 10.1038/sj.onc.1209009 PubMedGoogle Scholar
  27. 27.
    Luo J (2009) Glycogen synthase kinase 3beta (GSK3beta) in tumorigenesis and cancer chemotherapy. Cancer Lett 273:194–200. doi: 10.1016/j.canlet.2008.05.045 PubMedCrossRefGoogle Scholar
  28. 28.
    Gunes EG, Pinarbasi E, Pinarbasi H, Silig Y (2009) Strong association between lung cancer and the AXIN2 polymorphism. Mol Med Reports 2:1029–1035. doi: 10.3892/mmr_00000210 Google Scholar
  29. 29.
    Peterlongo P, Howe LR, Radice P, Sala P, Hong YJ et al (2005) Germline mutations of AXIN2 are not associated with nonsyndromic colorectal cancer. Hum Mutat 25:498–500. doi: 10.1002/humu.20189 PubMedCrossRefGoogle Scholar
  30. 30.
    Koch A, Weber N, Waha A, Hartmann W, Denkhaus D, Behrens J, Birchmeier W et al (2004) Mutations and elevated transcriptional activity of conductin (AXIN2) in hepatoblastomas. J Pathol 204:546–554. doi: 10.1002/path.1662 PubMedCrossRefGoogle Scholar
  31. 31.
    Koch A, Hrychyk A, Hartmann W, Waha A, Mikeska T, Waha A et al (2007) Mutations of the Wnt antagonist AXIN2 (Conductin) result in TCF-dependent transcription in medulloblastomas. Int J Cancer 121:284–291. doi: 10.1002/ijc.22675 PubMedCrossRefGoogle Scholar
  32. 32.
    Callahan N, Modesto A, Meira R, Seymen F, Patir A, Vieira AR (2009) Axis inhibition protein 2 (AXIN2) polymorphisms and tooth agenesis. Arch Oral Biol 54:45–49. doi: 10.1016/j.archoralbio.2008.08.002 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Fakhraddin Naghibalhossaini
    • 1
    • 2
    Email author
  • Mozhdeh Zamani
    • 1
  • Pooneh Mokarram
    • 1
  • Islam Khalili
    • 1
  • Mozhgan Rasti
    • 1
  • Zohreh Mostafavi-pour
    • 1
    • 3
  1. 1.Department of Biochemistry, School of MedicineShiraz University of Medical SciencesShirazIran
  2. 2.Autoimmune Research Center, School of MedicineShiraz University of Medical SciencesShirazIran
  3. 3.Recombinant Protein LaboratoryShiraz University of Medical SciencesShirazIran

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