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Downregulation of E-cadherin expression in breast cancer by promoter hypermethylation and its relation with progression and prognosis of tumor

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Abstract

Breast cancer is the most common cancer in women around the world, and novel prognosis strategies is needed to control more accurate and effective of this malignant disease. Among the latest prognostic markers is E-cadherin, which mediates cell–cell adhesion by associating with catenins. Loss of E-cadherin gene (CDH1) function by genetic or epigenetic alteration leads to tumorigenesis. The aim of our study was to investigate E-cadherin gene promoter methylation in breast cancer, and its correlation with E-cadherin protein expression. Fifty primary breast cancers tissue with ductal type and 50 normal breast sample from the same patients that was located adjacent to tumor region as controls were provided by Imam Reza-based referral and teaching hospital affiliated to Tabriz University of Medical Sciences, Tabriz, Iran. CDH1 promoter region CpG sites methylation and E-cadherin protein expression were determined by bisulfite-specific polymerase chain reaction and Western blot analysis, and the resulting products were sequenced on an ABI automated sequencer for firm conclusion. CDH1 hypermethylation in breast tumor specimen (ductal type) was observed in 94 % (47 of 50) comparing with normal samples methylation, and the significant difference was (p = 0.000). Protein expression in tumor samples tends to diminish with the CDH1 promoter region methylation. In the group of 50 ductal carcinomas cases, most of the cases showing CDH1 hypermethylation correlated inversely with the reduced levels of expression of E-cadherin proteins (95 % of full-methylated tumor samples had no protein expression, and 4.5 % of them had weak expression levels). Possible association was observed between CDH1 methylation and its protein expression (p = 0.000). The results of methylation analysis in promoter region in ten CpG sites (863, 865, 873, 879, 887, 892, 901, 918, 920, and 940) suggested that abnormal CDH1 methylation occurs in high frequencies in ductal breast tumors probably sounds the process of carcinogenesis progression.

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References

  1. Cervoni N, Szyf M. Demethylase activity is directed by histone acetylation. J Biol Chem. 2001;276:40778–87.

    Article  PubMed  CAS  Google Scholar 

  2. Chen ZX, Mann JR, Hsieh CL, Riggs AD, et al. Physical and functional interactions between the human DNMT3L protein and members of the de novo methyl transferase family. J Cell Biochem. 2005;95:902–17.

    Article  PubMed  CAS  Google Scholar 

  3. Herman JG, Baylin SB. Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med. 2003;349:2042–54.

    Article  PubMed  CAS  Google Scholar 

  4. Brown CJ. Role of the X chromosome in cancer. J Natl Cancer Inst. 1996;88:480–2.

    Article  PubMed  CAS  Google Scholar 

  5. Ribeiro-Filho LA, Franks J, Sasaki M, Shiina H, Li LC, Nojima D, et al. CpG hypermethylation of promoter region and inactivation of E-cadherin gene in human bladder cancer. Mol Carcinog. 2002;34(4):187–98.

    Article  PubMed  CAS  Google Scholar 

  6. Flanagan JM, Cocciardi S, Waddell N, Johnstone CN, et al. DNA methylome of familial breast cancer identifies distinct profiles defined by mutation status. Am J Hum Genet. 2010;86:420–33.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  7. Tan LW, Bianco T, Dobrovic A. Variable promotör region CpG island methylation of Putative tumor suppressor gene connexin 26 in breast cancer. Carcinogenesis. 2002;23:231–6.

    Article  PubMed  CAS  Google Scholar 

  8. Lee EYHP, Muller WJ. Oncogenes and tumor suppressor genes. Cold Spring Harb Perspect Biol. 2010;2:a003236.

    PubMed  CAS  PubMed Central  Google Scholar 

  9. Shen L, Kondo Y, Guo Y, Zhang J, Zhang L, Ahmed S, et al. Genome-wide profiling of DNA methylation reveals a class of normally methylated CpG island promoters. PLoS Genet. 2007;3(10):2023–36.

    Article  PubMed  CAS  Google Scholar 

  10. Rauch T, Pfeifer G. Methylated –CpG island recovery assay: a new technique for the rapid detection of methylated-CpG islands in cancer. Lab Investig. 2005;85:1172–80.

    Article  PubMed  CAS  Google Scholar 

  11. Nojima D, Nakajima K, Li LC, Franks J, Ribeiro-Filho L, Ishii N, et al. CpG methylation of promoter region inactivates E-cadherin gene in renal cell carcinoma. Mol Carcinog. 2001;32(1):19–27.

    Article  PubMed  CAS  Google Scholar 

  12. Sambrook J, Fritsch EF, Maniatis T. Separation of DNA in polyacrylamide gels. In: Molecular cloning: a laboratory manual. 2nd ed. New York: Cold Spring Harbor laboratory Press; 1989.p.125–30.

  13. Chao YL, Shepard CR, Wells A. Breast carcinoma cell re-expression E-cadherin during mesenchymal to epithelial reverting transition. Mol Cancer. 2010;9:179–97.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Brouckaert O, Schoneveld A, Truyers C, Kellen E, Van Ongeval C, Vergote I, et al. Breast cancer phenotype, nodal status and palpability may be useful in the detection of overdiagnosed screening-detected breast cancers. Ann Oncol. 2013;24(7):1847–52.

    Article  PubMed  CAS  Google Scholar 

  15. Balic M, Schwarzenbacher D, Stanzer S, Heitzer E, et al. Genetic and epigenetic analysis of putative breast cancer stem cell models. BMC Cancer. 2013;13:358.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  16. Matsumura T, Makino R, Mitamura K. Frequent down-regulation of E-cadherin by genetic and epigenetic changes in the malignant progression of hepatocellular carcinoma. Clin Cancer Res. 2001;7:594.

    PubMed  CAS  Google Scholar 

  17. Qiu X, Qiao F, Su X, Zhao Z, Fan H. Epigenetic activation of E-cadherin is a candidate therapeutic target in human hepatocellular carcinoma. Exp Ther Med. 2010;1(3):519–23.

    PubMed  CAS  PubMed Central  Google Scholar 

  18. Caldeira JR, Prando EC, Quevedo FC, Neto FA, Rainho CA, Rogatto SR. CDH1 promoter hypermethylation and E-cadherin protein expression in infiltrating breast cancer. BMC Cancer. 2006;6:48.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Sawada K, Mitra AK, Radjabi AR, Bhaskar V, et al. Loss of E-cadherin promotes ovarian cancer metastasis via α5- integrin, which is a therapeutic target. Cancer Res. 2008;68:2329–39.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  20. Lau MT, So WK, Leung PC. Fibroblast growth factor 2 induces E-cadherin down-regulation via PI3 K/Akt/mTOR and MAPK/ERK signaling in ovarian cancer cells.PLoS One. 2013;8(3):e59083.

  21. Liu YN, Lee WW, Wang CY, Chao TH, Chen Y, Chen JH. Regulatory mechanisms controlling human E-cadherin gene expression. Oncogene. 2005;24(56):8277–90.

    Article  PubMed  CAS  Google Scholar 

  22. Graziano F, Arduini F, Ruzzo A, Mandolesi A, et al. Combined analysis of E-cadherin gene (CDH1) promoter hypermethylation and E-cadherin protein expression in patients with gastric cancer: implications for treatment with demethylating drugs. Ann Oncol. 2004;15:489–92.

    Article  PubMed  CAS  Google Scholar 

  23. Walker DC, Southgate J. The modulatory effect of cell-cell contact on the tumourigenic potential of pre-malignant epithelial cells: a computational exploration. J R Soc Interface. 2013;10(78):20120703.

    Article  PubMed Central  Google Scholar 

  24. Fuchs M, Hermannstadter C, Specht K, Knyazev P, et al. Effect of tumor-associated mutant E-cadherin variant with defects in exons 8 or 9 on matrix metalloproteinase 3. J Cell Physiol. 2005;202:805–13.

    Article  PubMed  CAS  Google Scholar 

  25. Kleer CG, Van Golen KL, Braun T, Merajver SD. Persistant E-cadherin expression in inflammatory breast cancer. Mod Pathol. 2001;14:458–64.

    Article  PubMed  CAS  Google Scholar 

  26. Younis LK, El Sakka H, Haque I. The prognostic value of E-cadherin expression in breast cancer. Int J Health Sci (Qassim).2007;1(1):43–51.

  27. Kajabova V, Smolkova B, Zmetakova I, Sebova K, Krivulcik T, Bella V, et al. RASSF1A promoter methylation levels positively correlate with estrogen receptor expression in breast cancer patients. Transl Oncol. 2013;6(3):297–304.

    Article  PubMed  PubMed Central  Google Scholar 

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Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Medical Laboratory Science Department, Midwifery-Nursing Institute, Islamic Azad University of Chalous Branch, Chalous, Iran

    Shohreh Alizadeh Shargh

  2. Department of Medical Genetics and Biology, Health Institute of Eylul University, Izmir, Turkey

    Meral Sakizli

  3. Department of Biotechnology, Pasteur Institute of Iran, Tehran, Iran

    Vahid Khalaj

  4. Department of Medical Genetics, Pediatric Neurology Research Center, Mofid Hospital, Faculty of Medicine, Shahid Beheshti University of Medical Science, Chamran Highway, Velenjack St, Kodakyar Ave, Tehran, Islamic Republic of Iran

    Abolfazl Movafagh, Aresou Sayad & Neda Mansouri

  5. Department of Microbiology, Islamic Azad University of Varamin and Pishva Branch, Tehran, Iran

    Hamidreza Yazdi

  6. Department of Horticulture, Karaj Branch Islamic Azad University, Karaj, Iran

    Elmira Hagigatjou

  7. Proteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Seyed Abdolreza Mortazavi-Tabatabaei

  8. Genetic Research Centre, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran

    Hamid Reza Khorram Khorshid

Authors
  1. Shohreh Alizadeh Shargh
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  2. Meral Sakizli
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  3. Vahid Khalaj
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  4. Abolfazl Movafagh
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  6. Elmira Hagigatjou
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  8. Neda Mansouri
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  9. Seyed Abdolreza Mortazavi-Tabatabaei
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  10. Hamid Reza Khorram Khorshid
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Correspondence to Abolfazl Movafagh.

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Shargh, S.A., Sakizli, M., Khalaj, V. et al. Downregulation of E-cadherin expression in breast cancer by promoter hypermethylation and its relation with progression and prognosis of tumor. Med Oncol 31, 250 (2014). https://doi.org/10.1007/s12032-014-0250-y

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  • DOI: https://doi.org/10.1007/s12032-014-0250-y

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