Skip to main content
SpringerLink
Log in

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

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Malekzadeh R, Bishehsari F, Mahdavinia M, Ansari R (2009) Epidemiology and molecular genetics of colorectal cancer in Iran: a review. Arch Iran Med 12:161–169

    PubMed  CAS  Google Scholar 

  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

    PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  4. Angers S, Moon RT (2009) Proximal events in Wnt signal transduction. Natl Rev Mol Cell Biol 10:468–477. doi:10.1038/nrm2717

    CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  6. Behrens J, Lustig B (2004) The Wnt connection to tumorigenesis. Int J Dev Biol 48:477–487. doi:10.1387/ijdb.041815jb

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  Google Scholar 

  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)

    Article  PubMed  CAS  Google Scholar 

  9. Feinberg AP (2004) The epigenetics of cancer etiology. Semin Cancer Biol 14:427–432. doi:10.1016/j.semcancer.2004.06.005

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  12. Kikuchi A (1999) Modulation of Wnt signaling by axin and axil. Cytokine Growth Factor Rev 10:255–265

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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 18:279–284

    PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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 11:341–348

    PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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 60:4366–4371

    PubMed  CAS  Google Scholar 

  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

    Article  PubMed  Google Scholar 

  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

    Article  PubMed  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

  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

    Article  PubMed  CAS  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

  1. Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Zand Street, Shiraz, Iran

    Fakhraddin Naghibalhossaini, Mozhdeh Zamani, Pooneh Mokarram, Islam Khalili, Mozhgan Rasti & Zohreh Mostafavi-pour

  2. Autoimmune Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    Fakhraddin Naghibalhossaini

  3. Recombinant Protein Laboratory, Shiraz University of Medical Sciences, Shiraz, Iran

    Zohreh Mostafavi-pour

Authors
  1. Fakhraddin Naghibalhossaini
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mozhdeh Zamani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pooneh Mokarram
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Islam Khalili
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mozhgan Rasti
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zohreh Mostafavi-pour
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fakhraddin Naghibalhossaini.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Naghibalhossaini, F., Zamani, M., Mokarram, P. et al. Epigenetic and genetic analysis of WNT signaling pathway in sporadic colorectal cancer patients from Iran. Mol Biol Rep 39, 6171–6178 (2012). https://doi.org/10.1007/s11033-011-1434-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-011-1434-6

Keywords

Search

Navigation