Skip to main content

Advertisement

SpringerLink
Log in

High Frequency of Genes’ Promoter Methylation, but Lack of BRAF V600E Mutation among Iranian Colorectal Cancer Patients

  • Research
  • Published:
Pathology & Oncology Research

Abstract

Gene silencing due to DNA hypermethylation is a major mechanism for loss of tumor suppressor genes function in colorectal cancer. Activating V600E mutation in BRAF gene has been linked with widespread methylation of CpG islands in sporadic colorectal cancers. The aim of the present study was to evaluate the methylation status of three cancer-related genes, APC2, p14ARF, and ECAD in colorectal carcinogenesis and their association with the mutational status of BRAF and KRAS among Iranian colorectal cancer patients. DNA from 110 unselected series of sporadic colorectal cancer patients was examined for BRAF V600E mutation by PCR-RFLP. Promoter methylation of genes in tumors was determined by methylation specific PCR. The frequency of APC2, E-CAD, and p14 methylation was 92.6%, 40.4% and 16.7%, respectively. But, no V600E mutation was identified in the BRAF gene in any sample. No association was found in cases showing epigenetic APC, ECAD, and p14 abnormality with the clinicopathological parameters under study. The association between KRAS mutations and the so called methylator phenotype was previously reported. Therefore, we also analyzed the association between the hot spot KRAS gene mutations in codons of 12 and 13 with genes’ promoter hypermethylation in a subset of this group of patients. Out of 86 tumors, KRAS was mutated in 24 (28%) of tumors, the majority occurring in codon 12. KRAS mutations were not associated with genes’ methylation in this tumor series. These findings suggest a distinct molecular pathway for methylation of APC2, p14, and ECAD genes from those previously described for colorectal cancers with BRAF or KRAS mutations.

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

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. Fazeli MS, Adel MG, Lebaschi AH (2007) Colorectal carcinoma: a retrospective, descriptive study of age, gender, subsite, stage, and differentiation in Iran from 1995 to 2001 as observed in Tehran University. Dis Colon Rectum 50:990–995

    Article  PubMed  Google Scholar 

  3. Jass JR (2007) Classification of colorectal cancer based on correlation of clinical, morphological and molecular features. Histopathology 50:113e30

    Google Scholar 

  4. Toyota M, Ahuja N, Ohe-Toyota M, Herman JG, Baylin SB, Issa JP (1999) CpG island methylator phenotype in colorectal cancer. Proc Natl Acad Sci USA 96:8681–8686

    Article  PubMed  CAS  Google Scholar 

  5. Marais R, Marshall CJ (1996) Control of the ERK MAP kinase cascade by Ras and Raf. Cancer Surv 27:101–125

    PubMed  CAS  Google Scholar 

  6. Li WQ, Kawakami K, Ruszkiewicz A, Bennett G, Moore J, Iacopetta B (2006) BRAF mutations are associated with distinctive clinical, pathological and molecular features of colorectal cancer independently of microsatellite instability status. Mol Cancer 5:2

    Article  PubMed  Google Scholar 

  7. Davies H, Bignell GR, Cox C, Stephens P, Edkins S, Clegg S et al (2002) Mutations of the BRAF gene in human cancer. Nature 417:949–954

    Article  PubMed  CAS  Google Scholar 

  8. Rajagopalan H, Bardelli A, Lengauer C, Kinzler KW, Vogelstein B, Velculescu VE (2002) Tumorigenesis: RAF/RAS oncogenes and mismatch-repair status. Nature 418:934

    Article  PubMed  CAS  Google Scholar 

  9. Suehiro Y, Wong CW, Chirieac LR, Kondo Y, Shen L, Webb CR et al (2008) Epigenetic-genetic interactions in the APC/WNT, RAS/RAF, and P53 pathways in colorectal carcinoma. Clin Cancer Res 14:2560–2569

    Article  PubMed  CAS  Google Scholar 

  10. Weisenberger DJ, Siegmund KD, Campan M, Young J, Long TI, Faasse MA et al (2006) CpG island methylator phenotype underlies sporadic microsatellite instability and is tightly associated with BRAF mutation in colorectal cancer. Nat Genet 38:787–793

    Article  PubMed  CAS  Google Scholar 

  11. English DR, Young JP, Simpson JA, Jenkins MA, Southey MC, Walsh MD et al (2008) Ethnicity and risk for colorectal cancers showing somatic BRAF V600E mutation or CpG island methylator phenotype. Cancer Epidemiol Biomark Prev 17:1774–1780

    Article  CAS  Google Scholar 

  12. Nagasaka T, Sasamoto H, Notohara K, Cullings HM, Takeda M, Kimura K et al (2004) Colorectal cancer with mutation in BRAF, KRAS, and wild-type with respect to both oncogenes showing different patterns of DNA methylation. J Clin Oncol 22:4584–4594

    Article  PubMed  CAS  Google Scholar 

  13. Kominami K, Nagasaka T, Cullings HM, Hoshizima N, Sasamoto H, Young J et al (2009) Methylation in p14(ARF) is frequently observed in colorectal cancer with low-level microsatellite instability. J Int Med Res 37:1038–1045

    PubMed  CAS  Google Scholar 

  14. Auerkari EI (2006) Methylation of tumor suppressor genes p16 (INK4a), p27(Kip1) and E-cadherin in carcinogenesis. Oral Oncol 42:5–13

    Article  PubMed  CAS  Google Scholar 

  15. Schuebel KE, Chen W, Cope L, Glöckner SC, Suzuki H, Yi JM et al (2007) Comparing the DNA hypermethylome with gene mutations in human colorectal cancer. PLoS Genet 3:1709–1723

    Article  PubMed  CAS  Google Scholar 

  16. Kumar K, Brim H, Giardiello F, Smoot DT, Nouraie M, Lee EL et al (2009) Distinct BRAF (V600E) and KRAS mutations in high microsatellite instability sporadic colorectal cancer in African Americans. Clin Cancer Res 15:1155–1161

    Article  PubMed  CAS  Google Scholar 

  17. Garinis GA, Menounos PG, Spanakis NE, Papadopoulos K, Karavitis G, Parassi I et al (2002) Hypermethylation-associated transcriptional silencing of E-cadherin in primary sporadic colorectal carcinomas. J Pathol 198:442–449

    Article  PubMed  CAS  Google Scholar 

  18. Takeichi M (1995) Morphogenetic roles of classic cadherins. Curr Opin Cell Biol 7:619–627

    Article  PubMed  CAS  Google Scholar 

  19. Zheng S, Chen P, McMillan A, Lafuente A, Lafuente MJ, Ballesta A et al (2000) Correlations of partial and extensive methylation at the p14(ARF) locus with reduced mRNA expression in colorectal cancer cell lines and clinicopathological features in primary tumors. Carcinogenesis 21:2057–2064

    Article  PubMed  CAS  Google Scholar 

  20. 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

    Article  PubMed  CAS  Google Scholar 

  21. Servomaa K, Kiuru A, Kosma VM, Hirvikoski P, Rytömaa T (2000) p53 and K-ras gene mutations in carcinoma of the rectum among Finnish women. Mol Pathol 53:24–30

    Article  PubMed  CAS  Google Scholar 

  22. Gazin C, Wajapeyee N, Gobeil S, Virbasius CM, Green MR (2007) An elaborate pathway required for Ras-mediated epigenetic silencing. Nature 449:1073–1077

    Article  PubMed  CAS  Google Scholar 

  23. Mokarram P, Kumar K, Brim H, Naghibalhossaini F, Saberi-firoozi M, Nouraie M et al (2009) Distinct high-profile methylated genes in colorectal cancer. PLoS ONE 4:e7012

    Article  PubMed  Google Scholar 

  24. Miranda E, Destro A, Malesci A, Balladore E, Bianchi P, Baryshnikova E et al (2006) Genetic and epigenetic changes in primary metastatic and nonmetastatic colorectal cancer. Br J Cancer 95:1101–1107

    Article  PubMed  CAS  Google Scholar 

  25. Wheeler JM, Kim HC, Efstathiou JA, Ilyas M, Mortensen NJ, Bodmer WF (2001) Hypermethylation of the promoter region of the E-cadherin gene (CDH1) in sporadic and ulcerative colitis associated colorectal cancer. Gut 48:367–371

    Article  PubMed  CAS  Google Scholar 

  26. Strathdee G (2002) Epigenetic versus genetic alterations in the inactivation of E-cadherin. Semin Cancer Biol 12:373–379

    Article  PubMed  CAS  Google Scholar 

  27. van Engeland M, Weijenberg MP, Roemen GM, Brink M, de Bruïne AP, Goldbohm RA et al (2003) Effects of dietary folate and alcohol intake on promoter methylation in sporadic colorectal cancer: the Netherlands cohort study on diet and cancer. Cancer Res 63:3133–3137

    PubMed  Google Scholar 

  28. 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

    Article  PubMed  CAS  Google Scholar 

  29. Yuen ST, Davies H, Chan TL, Ho JW, Bignell GR, Cox C et al (2002) Similarity of the phenotypic patterns associated with BRAF and KRAS mutations in colorectal neoplasia. Cancer Res 62:6451–6455

    PubMed  CAS  Google Scholar 

  30. Ogino S, Kawasaki T, Kirkner GJ, Kraft P, Loda M, Fuchs CS (2007) Evaluation of markers for CpG island methylator phenotype (CIMP) in colorectal cancer by a large population-based sample. J Mol Diagn 9:305–314

    Article  PubMed  CAS  Google Scholar 

  31. Brim H, Mokarram P, Naghibalhossaini F, Saberi-Firoozi M, Al-Mandhari M, Al-Mawaly K et al (2008) Impact of BRAF, MLH1 on the incidence of microsatellite instability high colorectal cancer in populations based study. Mol Cancer 7:68

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Dr. Mahmood Vessal of Shiraz University of Medical Sciences for carefully reading the manuscript. This study was part of the dissertation of Hamideh Mahmoodzadeh Hosseinini, submitted to Shiraz University of Medical Sciences in partial fulfillment of the requirements for the MSc 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, Shiraz University of Medical Sciences, School of Medicine, Zand Street, Shiraz, Fars, Iran

    Fakhraddin Naghibalhossaini, Hamideh Mahmoodzadeh Hosseini, Pooneh Mokarram & Mozhdeh Zamani

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

    Fakhraddin Naghibalhossaini

  3. Gastroenterohepathology Research Centre, Shiraz University of Medical Sciences, Shiraz, Iran

    Pooneh Mokarram

Authors
  1. Fakhraddin Naghibalhossaini
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hamideh Mahmoodzadeh Hosseini
    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. Mozhdeh Zamani
    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., Hosseini, H.M., Mokarram, P. et al. High Frequency of Genes’ Promoter Methylation, but Lack of BRAF V600E Mutation among Iranian Colorectal Cancer Patients. Pathol. Oncol. Res. 17, 819–825 (2011). https://doi.org/10.1007/s12253-011-9388-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12253-011-9388-5

Keywords

Search

Navigation