Annals of Laboratory Medicine

Korean Society for Laboratory Medicine (대한진단검사의학회)
권호 표지
DOI QR코드

DOI QR Code

Comparison of Methylation Profiling in Cancerous and Their Corresponding Normal Tissues from Korean Patients with Breast Cancer

  • Jung, Eun-Jung (Division of Surgical Oncology, Department of Surgery, Gyeongsang National University Hospital) ;
  • Kim, In-Suk (Department of Laboratory Medicine, Biomedical Research Institute, Pusan National University Hospital) ;
  • Lee, Eun Yup (Department of Laboratory Medicine, Biomedical Research Institute, Pusan National University Hospital) ;
  • Kang, Jeong-Eun (Department of Laboratory Medicine, Pusan National University Yangsan Hospital) ;
  • Lee, Sun-Min (Department of Laboratory Medicine, Pusan National University Yangsan Hospital) ;
  • Kim, Dong Chul (Department of Pathology, Gyeongsang National University Hospital) ;
  • Kim, Ju-Yeon (Division of Surgical Oncology, Department of Surgery, Gyeongsang National University Hospital) ;
  • Park, Soon-Tae (Division of Surgical Oncology, Department of Surgery, Gyeongsang National University Hospital)
  • Published : 2013.11.01
⟨ Previous Next ⟩

Abstract

Background: Aberrant DNA hypermethylation plays a pivotal role in carcinogenesis and disease progression; therefore, accurate measurement of differential gene methylation patterns among many genes is likely to reveal biomarkers for improved risk assessment. We evaluated the gene hypermethylation profiles of primary breast tumors and their corresponding normal tissues and investigated the association between major clinicopathological features and gene hypermethylation. Methods: A single reaction using methylation-specific multiplex ligation-dependent probe amplification was used to analyze the DNA methylation status of 24 tumor suppressor genes in 60 cancerous tissues and their corresponding normal tissues from patients with primary breast cancer. Results: In cancerous breast tissues, 21 of 24 genes displayed promoter methylation in one or more samples. The most frequently methylated genes included RASSF1 (43.3%), APC (31.7%), CDKN2B (25.0%), CDH13 (23.3%), GSTP1 (16.7%), and BRCA1 (10%). APC was associated with lymph node metastasis, and BRCA1 was associated with negative estrogen receptor and negative progesterone receptor expression. In normal breast tissues, 8 of 24 tumor suppressor genes displayed promoter hypermethylation; CDKN2B (28.3%) and RASSF1 (8.3%) hypermethylation were most frequently observed. Conclusions: RASSF1 and CDKN2B hypermethylation in Korean breast cancer patients were the most frequent in cancerous tissue and corresponding normal tissue, respectively. Our data indicates that methylation of specific genes is a frequent event in morphologically normal breast tissues adjacent to breast tumors as well as the corresponding breast cancers. This study also suggests that gene methylation is linked to various pathological features of breast cancer; however, this requires confirmation in a larger study.

Keywords

References

  1. The Korea Central Cancer Registry. Annual report of cancer statistics in Korea in 2009. Seoul: The Korean Ministry of Health and Welfare, 2011.
  2. Li S, Rong M, Iacopetta B. DNA hypermethylation in breast cancer and its association with clinicopathological features. Cancer Lett 2006;237:272-80. https://doi.org/10.1016/j.canlet.2005.06.011
  3. Van De Voorde L, Speeckaert R, Van Gestel D, Bracke M, De Neve W, Delanghe J, et al. DNA methylation-based biomarkers in serum of patients with breast cancer. Mutat Res 2012;751:304-25. https://doi.org/10.1016/j.mrrev.2012.06.001
  4. Marzese DM, Hoon DS, Chong KK, Gago FE, Orozco JI, Tello OM, et al. DNA methylation index and methylation profile of invasive ductal breast tumors. J Mol Diagn 2012;14:613-22. https://doi.org/10.1016/j.jmoldx.2012.07.001
  5. Cheol Kim D, Thorat MA, Lee MR, Cho SH, Vasiljevíc N, Scibior-Bentkowska D, et al. Quantitative DNA methylation and recurrence of breast cancer: a study of 30 candidate genes. Cancer Biomark 2012;11:75-88. https://doi.org/10.3233/CBM-2012-0266
  6. Zhu W, Qin W, Hewett JE, Sauter ER. Quantitative evaluation of DNA hypermethylation in malignant and benign breast tissue and fluids. Int J Cancer 2010;126:474-82. https://doi.org/10.1002/ijc.24728
  7. Esteller M. CpG island hypermethylation and tumor suppressor genes: a booming present, a brighter future. Oncogene 2002;21:5427-40. https://doi.org/10.1038/sj.onc.1205600
  8. Esteller M. Cancer epigenetics: DNA methylation and chromatin alterations in human cancer. Adv Exp Med Biol 2003;532:39-49. https://doi.org/10.1007/978-1-4615-0081-0_5
  9. Widschwendter M and Jones PA. DNA methylation and breast carcinogenesis. Oncogene 2002;21:5462-82. https://doi.org/10.1038/sj.onc.1205606
  10. Xu J, Shetty PB, Feng W, Chenault C, Bast RC Jr, Issa JP, et al. Methylation of HIN-1, RASSF1A, RIL and CDH13 in breast cancer is associated with clinical characteristics, but only RASSF1A methylation is associated with outcome. BMC Cancer 2012;12:243. https://doi.org/10.1186/1471-2407-12-243
  11. Wang S, Dorsey TH, Terunuma A, Kittles RA, Ambs S, Kwabi-Addo B. Relationship between tumor DNA methylation status and patient characteristics in African-American and European-American women with breast cancer. PLoS One 2012;7:e37928. https://doi.org/10.1371/journal.pone.0037928
  12. van Hoesel AQ, van de Velde CJ, Kuppen PJ, Putter H, de Kruijf EM, van Nes JG, et al. Primary tumor classification according to methylation pattern is prognostic in patients with early stage ER-negative breast cancer. Breast Cancer Res Treat 2012;131:859-69. https://doi.org/10.1007/s10549-011-1485-3
  13. Tserga A, Michalopoulos NV, Levidou G, Korkolopoulou P, Zografos G, Patsouris E, et al. Association of aberrant DNA methylation with clinicopathological features in breast cancer. Oncol Rep 2012;27:1630-8.
  14. Szyf M. DNA methylation signatures for breast cancer classification and prognosis. Genome Med 2012;4:26. https://doi.org/10.1186/gm325
  15. Elston CW and Ellis IO. Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long-term follow-up. Histopathology 2002;41:154-61. https://doi.org/10.1046/j.1365-2559.2002.14892.x
  16. Wolff AC, Hammond ME, Schwartz JN, Hagerty KL, Allred DC, Cote RJ, et al. American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. J Clin Oncol 2007;25:118-45.
  17. Nygren AO, Ameziane N, Duarte HM, Vijzelaar RN, Waisfisz Q, Hess CJ, et al. Methylation-specific MLPA (MS-MLPA): simultaneous detection of CpG methylation and copy number changes of up to 40 sequences. Nucleic Acids Res 2005;33:e128. https://doi.org/10.1093/nar/gni127
  18. Van der Auwera I, Bovie C, Svensson C, Trinh XB, Limame R, van Dam P, et al. Quantitative methylation profiling in tumor and matched morphologically normal tissues from breast cancer patients. BMC Cancer 2010;10:97. https://doi.org/10.1186/1471-2407-10-97
  19. Park SY, Kwon HJ, Lee HE, Ryu HS, Kim SW, Kim JH, et al. Promoter CpG island hypermethylation during breast cancer progression. Virchows Arch 2011;458:73-84. https://doi.org/10.1007/s00428-010-1013-6
  20. Kim JH, Shin MH, Kweon SS, Park MH, Yoon JH, Lee JS, et al. Evaluation of promoter hypermethylation detection in serum as a diagnostic tool for breast carcinoma in Korean women. Gynecol Oncol 2010;118:176-81. https://doi.org/10.1016/j.ygyno.2010.04.016
  21. Marzese DM, Gago FE, Vargas-Roig LM, Roque M. Simultaneous analysis of the methylation profile of 26 cancer related regions in invasive breast carcinomas by MS-MLPA and drMS-MLPA. Mol Cell Probes 2010;24:271-80. https://doi.org/10.1016/j.mcp.2010.05.002
  22. Buyru N, Altinisik J, Ozdemir F, Demokan S, Dalay N. Methylation profiles in breast cancer. Cancer Invest 2009;27:307-12. https://doi.org/10.1080/07357900802350814
  23. Marzese DM, Gago FE, Vargas-Roig LM, Roque M. Simultaneous analysis of the methylation profile of 26 cancer related regions in invasive breast carcinomas by MS-MLPA and drMS-MLPA. Mol Cell Probes 2010;24:271-80. https://doi.org/10.1016/j.mcp.2010.05.002
  24. Dammann R, Yang G, Pfeifer GP. Hypermethylation of the cpG island of Ras association domain family 1A (RASSF1A), a putative tumor suppressor gene from the 3p21.3 locus, occurs in a large percentage of human breast cancers. Cancer Res 2001;61:3105-9.
  25. Honorio S, Agathanggelou A, Schuermann M, Pankow W, Viacava P, Maher ER, et al. Detection of RASSF1A aberrant promoter hypermethylation in sputum from chronic smokers and ductal carcinoma in situ from breast cancer patients. Oncogene 2003;22:147-50. https://doi.org/10.1038/sj.onc.1206057
  26. Yeo W, Wong WL, Wong N, Law BK, Tse GM, Zhong S. High frequency of promoter hypermethylation of RASSF1A in tumorous and non-tumourous tissue of breast cancer. Pathology 2005;37:125-30. https://doi.org/10.1080/00313020500058623
  27. Muller HM, Widschwendter A, Fiegl H, Ivarsson L, Goebel G, Perkmann E, et al. DNA methylation in serum of breast cancer patients: an independent prognostic marker. Cancer Res 2003;63:7641-5.
  28. Mehrotra J, Vali M, McVeigh M, Kominsky SL, Fackler MJ, Lahti-Domenici J, et al. Very high frequency of hypermethylated genes in breast cancer metastasis to the bone, brain, and lung. Clin Cancer Res 2004;10:3104-9. https://doi.org/10.1158/1078-0432.CCR-03-0118
  29. Dodge JE, Munson C, List AF. KG-1 and KG-1a model the p15 CpG island methylation observed in acute myeloid leukemia patients. Leuk Res 2001;25:917-25. https://doi.org/10.1016/S0145-2126(01)00053-4
  30. Aggerholm A, Guldberg P, Hokland M, Hokland P. Extensive intra- and interindividual heterogeneity of p15INK4B methylation in acute myeloid leukemia. Cancer Res 1999;59:436-41.
  31. Herman JG, Jen J, Merlo A, Baylin SB. Hypermethylation-associated inactivation indicates a tumor suppressor role for p15INK4B. Cancer Res 1996;56:722-7.
  32. Christiansen DH, Andersen MK, Pedersen-Bjergaard J. Methylation of p15INK4B is common, is associated with deletion of genes on chromosome arm 7q and predicts a poor prognosis in therapy-related myelodysplasia and acute myeloid leukemia. Leukemia 2003;17:1813-9. https://doi.org/10.1038/sj.leu.2403054
  33. Qin Y, Liu JY, Li B, Sun ZL, Sun ZF. Association of low p16INK4a and p15INK4b mRNAs expression with their CpG islands methylation with human hepatocellular carcinogenesis. World J Gastroenterol 2004;10:1276-80. https://doi.org/10.3748/wjg.v10.i9.1276
  34. Ishiguro A, Takahata T, Saito M, Yoshiya G, Tamura Y, Sasaki M, et al. Influence of methylated p15 and p16 genes on clinicopathological features in colorectal cancer. J Gastroenterol Hepatol 2006;21:1334-9. https://doi.org/10.1111/j.1440-1746.2006.04137.x
  35. Jin Z, Tamura G, Tsuchiya T, Sakata K, Kashiwaba M, Osakabe M, et al. Adenomatous polyposis coli (APC) gene promoter hypermethylation in primary breast cancers. Br J Cancer 2001;85:69-73. https://doi.org/10.1054/bjoc.2001.1853

Cited by

  1. The functional role of long non-coding RNAs and epigenetics vol.16, pp.None, 2013, https://doi.org/10.1186/1480-9222-16-11
  2. Epigenetic mechanisms of breast cancer: an update of the current knowledge vol.6, pp.6, 2013, https://doi.org/10.2217/epi.14.59
  3. Genetic risk variants associated with in situ breast cancer vol.17, pp.None, 2015, https://doi.org/10.1186/s13058-015-0596-x
  4. Association of BRCA1 promoter methylation with sporadic breast cancers: Evidence from 40 studies vol.5, pp.None, 2013, https://doi.org/10.1038/srep17869
  5. Increased Hypermethylation of Glutathione S-Transferase P1, DNA-Binding Protein Inhibitor, Death Associated Protein Kinase and Paired Box Protein-5 Genes in Triple-Negative Breast Cancer Saudi Females vol.16, pp.2, 2013, https://doi.org/10.7314/apjcp.2015.16.2.541
  6. Epigenetic markers in melanoma vol.2, pp.4, 2015, https://doi.org/10.2217/mmt.15.30
  7. Exploring DNA methylation changes in promoter, intragenic, and intergenic regions as early and late events in breast cancer formation vol.15, pp.None, 2013, https://doi.org/10.1186/s12885-015-1777-9
  8. Aberrant GSTP1 promoter methylation is associated with increased risk and advanced stage of breast cancer: a meta-analysis of 19 case-control studies vol.15, pp.None, 2015, https://doi.org/10.1186/s12885-015-1926-1
  9. Promoter methylation of TRIM9 as a marker for detection of circulating tumor DNA in breast cancer patients vol.4, pp.1, 2013, https://doi.org/10.1186/s40064-015-1423-7
  10. Quantitative assessment of the association between APC promoter methylation and breast cancer vol.7, pp.25, 2013, https://doi.org/10.18632/oncotarget.9354
  11. Association between aberrant APC promoter methylation and breast cancer pathogenesis: a meta-analysis of 35 observational studies vol.4, pp.None, 2016, https://doi.org/10.7717/peerj.2203
  12. Genomic Changes in Normal Breast Tissue in Women at Normal Risk or at High Risk for Breast Cancer vol.10, pp.None, 2013, https://doi.org/10.4137/bcbcr.s39384
  13. A Systematic Analysis of the Relationship of CDH13 Promoter Methylation and Breast Cancer Risk and Prognosis vol.11, pp.5, 2013, https://doi.org/10.1371/journal.pone.0149185
  14. Recent Progress in Triple Negative Breast Cancer Research vol.17, pp.4, 2013, https://doi.org/10.7314/apjcp.2016.17.4.1595
  15. Methylation Profiles of BRCA1, RASSF1A and GSTP1 in Vietnamese Women with Breast Cancer vol.19, pp.7, 2013, https://doi.org/10.22034/apjcp.2018.19.7.1887
  16. Promoter hypermethylation of RARβ2, DAPK, hMLH1, p14, and p15 is associated with progression of breast cancer : A PRISMA-compliant meta-analysis vol.97, pp.51, 2018, https://doi.org/10.1097/md.0000000000013666
  17. Epigenetic modulation of BRCA‐1 and MGMT genes, and histones H4 and H3 are associated with breast tumors vol.120, pp.8, 2013, https://doi.org/10.1002/jcb.28645
  18. The association between the methylation frequency of BRCA1/2 gene promoter and occurrence and prognosis of breast carcinoma : A meta-analysis vol.99, pp.10, 2013, https://doi.org/10.1097/md.0000000000019345
  19. The Clinical and Pathological Profile of BRCA1 Gene Methylated Breast Cancer Women: A Meta-Analysis vol.13, pp.6, 2013, https://doi.org/10.3390/cancers13061391