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Tumor Biology

, Volume 31, Issue 4, pp 321–331 | Cite as

CpG island methylator phenotype of multigene in serum of sporadic breast carcinoma

  • Feng Jing
  • Wang Yuping
  • Chen Yong
  • Luo Jie
  • Lu Jun
  • Tang Xuanbing
  • Hu Lihua
Research Article

Abstract

CpG island methylator phenotype (CIMP) involves methylation targeted toward the promoters of multiple genes. We determined a methylation profile of tumor-related genes in serum of sporadic breast cancer (SBC). The multigene methylation was examined by methylation-specific polymerase chain reaction assay in serum of 50 SBCs and 50 paired nontumors, and CIMP+ was defined as having three genes that are concordantly methylated. The methylation frequency of ten genes in serum of 50 SBCs varied from 10% in FHIT to 74% in RASSF1A. The methylation status of RASSF1A, BRCA1, p16, CDH1, ER, RARβ2, APC, and DAPK was significantly correlated with SBC and nontumor serum (P < 0.05). Methylation of at least one gene was found in 92% SBC; CIMP was more frequent in SBC than nontumor serum (P < 0.001). There was a significant association between CIMP and methylation of RASSF1A, BRCA1, p16, CDH1, ER, RARβ2, APC, and DAPK (P < 0.05); the methylation link profile of CDH1, RASSF1A, BRCA1, and RARβ2 as breast cancer marker may contribute high sensitivity (90%) and specificity (88%). ER and RARβ2 methylation was associated with elevated serum CA153 levels in 39 SBC samples with CIMP+ (P < 0.05). Multivariate analysis showed that living area of patients was found to provide independent prognostic information associated with a relative risk of tumor recurrence of 5.3. Multigene-specific methylation profile in serum was association with the recurrence risk of rural SBC, and positive correlation of CIMP can serve as a promising molecular marker of SBC.

Keywords

CpG island methylator phenotype Sporadic breast cancer Methylation profile 

Notes

Grants provided by Science Funds for Distinguished Young Scholar of Hubei Province (2008154), Natural Science Foundation of Hubei Province (2007ABA371), Science Funds from Hubei Provincial Department of Education (Q20082407), and Innovation Foundation for Young Scholars of Yunyang Medical College (CXX200805).

References

  1. 1.
    Goldhirsch A, Glick JH, Gelber RD, et al. Meeting highlights: international consensus panel on the treatment of primary breast cancer. Seventh international conference on adjuvant therapy of primary breast cancer. J Clin Oncol. 2001;19:3817–27.PubMedGoogle Scholar
  2. 2.
    Early Breast Cancer Trialists' Collaborative Group. Polychemotherapy for early breast cancer: an overview of the randomised trials. Early breast cancer trialists' collaborative group. Lancet. 1998;352:930–42.CrossRefGoogle Scholar
  3. 3.
    Early Breast Cancer Trialists' Collaborative Group. Tamoxifen for early breast cancer: an overview of the randomised trials. Early breast cancer trialists' collaborative group. Lancet. 1998;351:1451–67.CrossRefGoogle Scholar
  4. 4.
    Hayes DF, Isaacs C, Stearns V. Prognostic factors in breast cancer: current and new predictors of metastasis. J Mammary Gland Biol Neoplasia. 2001;6:375–92.CrossRefPubMedGoogle Scholar
  5. 5.
    Couch FJ, Weber BL. Breast cancer. In: Scriver CR, Beaudet AL, Sly WS, et al., editors. The metabolic and molecular basis of human disease. New York: McGraw-Hill; 2001. p. 999–1031.Google Scholar
  6. 6.
    Leitch AM. Breast cancer screening: success amid conflict. Surg Oncol Clin N Am. 1999;8:657–72.PubMedGoogle Scholar
  7. 7.
    Dulaimi E, Hillinck J, Ibanez CI, et al. Tumor suppressor gene promoter hypermethylation in serum of breast cancer patients. Clin Cancer Res. 2004;15(10):6189–93.CrossRefGoogle Scholar
  8. 8.
    Müller HM, Fiegl H, Widschwendter A, et al. Prognostic DNA methylation marker in serum of cancer patients. Ann N Y Acad Sci. 2004;1022:44–9.CrossRefPubMedGoogle Scholar
  9. 9.
    Hu XC, Wong IH, Chow LW. Tumor-derived aberrant methylation in plasma of invasive ductal breast cancer patients: clinical implications. Oncol Rep. 2003;10(6):1811–5.PubMedGoogle Scholar
  10. 10.
    Umbricht CB, Evron E, Gabrielson E, et al. Hypermethylation of 14-3-3 sigma (stratifin) is an early event in breast cancer. Oncogene. 2001;20:3348–53.CrossRefPubMedGoogle Scholar
  11. 11.
    Evron E, Dooley WC, Umbricht CB, et al. Detection of breast cancer cells in ductal lavage fluid by methylation-specific PCR. Lancet. 2001;357:1335–6.CrossRefPubMedGoogle Scholar
  12. 12.
    Evron E, Umbricht CB, Korz D, et al. Loss of cyclin D2 expression in the majority of breast cancers is associated with promoter hypermethylation. Cancer Res. 2001;61:2782–7.PubMedGoogle Scholar
  13. 13.
    Ulivi P, Zoli W, Calistri D, et al. p16INK4A and CDH13 hypermethylation in tumor and serum of non-small cell lung cancer patients. J Cell Physiol. 2006;206(3):611–5.CrossRefPubMedGoogle Scholar
  14. 14.
    Belinsky SA, Nikula KJ, Palmisano WA, et al. Aberrant methylation of p16(INK4a) is an early event in lung cancer and a potential biomarker for early diagnosis. PNAS. 1998;95:11891–6.CrossRefPubMedGoogle Scholar
  15. 15.
    Tsou JA, Hagen JA, Carpenter CL. DNA methylation analysis: a powerful new tool for lung cancer diagnosis. Oncogene. 2002;21(35):5450–61.CrossRefPubMedGoogle Scholar
  16. 16.
    Usadel H, Brabender J, Danenberg KD. Quantitative adenomatous polyposis coli promoter methylation analysis in tumor tissue, serum, and plasma DNA of patients with lung cancer. Cancer Res. 2002;62(2):371–5.PubMedGoogle Scholar
  17. 17.
    Barault L, Charon-Barra C, Jooste V. Hypermethylator phenotype in sporadic colon cancer: study on a population-based series of 582 cases. Cancer Res. 2008;68(20):8541–6.CrossRefPubMedGoogle Scholar
  18. 18.
    Esteller M, Sparks A, Toyota M, et al. Analysis of adenomatous polyposis coli promoter hypermethylation in human cancer. Cancer Res. 2000;60:4366–71.PubMedGoogle Scholar
  19. 19.
    Ngan RK, Lau WH, Yip TT, et al. Remarkable application of serum EBV EBER-1 in monitoring response of nasopharyngeal cancer patients to salvage chemotherapy. Ann N Y Acad Sci. 2001;945:73–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Lo YM. Prognostic implication of pretreatment plasma/serum concentration of Epstein-Barr virus DNA in nasopharyngeal carcinoma. Biomed Pharmacother. 2001;55:362–5.CrossRefPubMedGoogle Scholar
  21. 21.
    Sidransky D, Von Eschenbach A, Tsai YC, et al. Identification of p53 gene mutations in bladder cancers and urine samples. Science. 1991;252:706–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Muller HM, Widschwendter A, Fiegl H, et al. DNA methylation in serum of breast cancer patients: an independent prognostic marker. Cancer Res. 2003;63:7641–5.PubMedGoogle Scholar
  23. 23.
    Jeronimo C, Costa I, Martins MC, et al. Detection of gene promoter hypermethylation in fine needle washings from breast lesions. Clin Cancer Res. 2003;9:3413–7.PubMedGoogle Scholar
  24. 24.
    Hoque MO, Begum S, Topaloglu O, Jeronimo C, et al. Quantitative detection of promoter hypermethylation of multiple genes in the tumor, urine, and serum DNA of patients with renal cancer. Cancer Res. 2004;64:5511–17.CrossRefPubMedGoogle Scholar
  25. 25.
    Kawakami K, Brabender J, Lord RV, et al. Hypermethylated APC DNA in plasma and prognosis of patients with esophageal adenocarcinoma. J Natl Cancer Inst. 2000;92:1805–11.CrossRefPubMedGoogle Scholar
  26. 26.
    Topaloglu O, Hoque MO, Tokumaru Y, et al. Detection of promoter hypermethylation of multiple genes in the tumor and bronchoalveolar lavage of patients with lung cancer. Clin Cancer Res. 2004;10:2284–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Yang HJ, Liu VW, Wang Y, et al. Detection of hypermethylated genes in tumor and plasma of cervical cancer patients. Gynecol Oncol. 2004;93:435–40.CrossRefPubMedGoogle Scholar
  28. 28.
    Usadel H, Brabender J, Danenberg KD, et al. Quantitative adenomatous polyposis coli promoter methylation analysis in tumor tissue, serum, and plasma DNA of patients with lung cancer. Cancer Res. 2002;62:371–5.PubMedGoogle Scholar
  29. 29.
    Toyota M, Ahuja N, Ohe-Toyota M, et al. CpG island methylator phenotype in colorectal cancer. Proc Natl Acad Sci USA. 1999;96(15):8681–6.CrossRefPubMedGoogle Scholar
  30. 30.
    Ogino S, Cantor M, Kawasaki T, et al. CpG island methylator phenotype (CIMP) of colorectal cancer is best characterised by quantitative DNA methylation analysis and prospective cohort studies. Gut. 2006;55(7):1000–6.CrossRefPubMedGoogle Scholar
  31. 31.
    Zhang C, Li Z, Cheng Y, et al. CpG island methylator phenotype association with elevated serum alpha-fetoprotein level in hepatocellular carcinoma. Clin Cancer Res. 2007;13(3):944–52.CrossRefPubMedGoogle Scholar
  32. 32.
    Toyota M, Ahuja N, Ohe-Toyota M, Herman JG, Baylin SB, Issa JP. CpG island methylator phenotype in colorectal cancer. Proc Natl Acad Sci USA. 1999;96:8681–6.CrossRefPubMedGoogle Scholar
  33. 33.
    Toyota M, Ahuja N, Suzuki H, et al. Aberrant methylation in gastric cancer associated with the CpG island methylator phenotype. Cancer Res. 1999;59:5438–42.PubMedGoogle Scholar
  34. 34.
    Weisenberger DJ, Siegmund KD, Campan M, et al. CpG island methylator phenotype underlies sporadic microsatellite instability and is tightly associated with BRAF mutation in colorectal cancer. Nat Genet. 2006;38:787–93.CrossRefPubMedGoogle Scholar
  35. 35.
    Sambrook J, Russell DW. Molecular cloning. A laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 2001.Google Scholar
  36. 36.
    Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA. 1996;93:9821–6.CrossRefPubMedGoogle Scholar
  37. 37.
    Grunau C, Clark SJ, Rosenthal A. Bisulifite genomic sequencing: systematic investigation of critical experimental parameters. Nucleic Acids Res. 2001;29:e65.CrossRefPubMedGoogle Scholar
  38. 38.
    Esteller M, Silva JM, Dominguez G, Bonilla F, Xavier MG, Lerma E, et al. Promoter hypermethylation and BRCA1 inactivation in sporadic breast and ovarian tumors. J Natl Cancer Inst (Bethesda). 2000;92:564–9.CrossRefGoogle Scholar
  39. 39.
    Sabine ZM, Fong KM, Maitra A, et al. 50 CpG island methylation of the FHIT gene is correlated with loss of gene expression in lung and breast cancer. Cancer Res. 2001;61:3581–5.Google Scholar
  40. 40.
    Katzenellenbogen RA, Baylin SB, Herman JG. Hypermethylation of the DAP-kinase CpG island is a common alteration in B-cell malignancies. Blood. 1999;93:4347–53.PubMedGoogle Scholar
  41. 41.
    Burbee DG, Forgacs E, Zochbauer-Muller S, et al. Epigenetic inactivation of RASSF1A in lung and breast cancers and malignant phenotype suppression. J Natl Cancer Inst. 2001;93:691–9.CrossRefPubMedGoogle Scholar
  42. 42.
    Widschwendter M, Berger J, Hermann M, et al. Methylation and silencing of the retinoic acid receptor-beta2 gene in breast cancer. J Natl Cancer Inst. 2001;92:826–32.CrossRefGoogle Scholar
  43. 43.
    Corn PG, Smith BD, Ruckdeschel ES, Douglas D, Baylin SB, Herman JG. E-cadherin expression is silenced by 5 CpG island methylation in acute leukemia. Clin Cancer Res. 2000;6:4243–8.PubMedGoogle Scholar
  44. 44.
    Mahmood A, Yulan C, Magno RM. Promoter methylation regulates helicobacter pylori-stimulated cyclooxygenase-2 expression in gastric epithelial cells. Cancer Res. 2001;61:2399–403.Google Scholar
  45. 45.
    Mori T, Martinez SR, O'Day SJ. Estrogen receptor-α methylation predicts melanoma progression. Cancer Res. 2006;66:6692–8.CrossRefPubMedGoogle Scholar
  46. 46.
    Jones PA, Takai D. The role of DNA methylation in mammalian epigenetics. Science (Wash DC). 2001;293:1068–70.CrossRefGoogle Scholar
  47. 47.
    Maike W, Andreas H, Andrea B, et al. Methylation of serum DNA is an independent prognostic marker in colorectal cancer. Clin Cancer Res. 2006;12:7347–52.CrossRefGoogle Scholar
  48. 48.
    Misawa K, Ueda Y, Kanazawa T, et al. Epigenetic inactivation of galanin receptor 1 in head and neck cancer. Clin Cancer Res. 2008;14:7604–13.CrossRefPubMedGoogle Scholar
  49. 49.
    Euhus DM, Bu D, Milchgrub S, et al. DNA methylation in benign breast epithelium in relation to age and breast cancer risk. Cancer Epidemiol Biomark Prev. 2008;17:1051–9.CrossRefGoogle Scholar
  50. 50.
    Anders R, Jörg T, Solvang HK, et al. GSTP1 promoter haplotypes affect DNA methylation levels and promoter activity in breast carcinomas. Cancer Res. 2008;68:5562–71.CrossRefGoogle Scholar
  51. 51.
    Oue N, Mitani Y, Motoshita J, et al. Accumulation of DNA methylation is associated with tumor stage in gastric cancer. Cancer. 2006;106:1250–9.CrossRefPubMedGoogle Scholar
  52. 52.
    Eads CA, Lord RV, Wickramasinghe K, et al. Epigenetic patterns in the progression of esophageal adenocarcinoma. Cancer Res. 2001;61:3410–8.PubMedGoogle Scholar
  53. 53.
    Yamashita K, Dai T, Dai Y, Yamamoto F, Perucho M. Genetics supersedes epigenetics in colon cancer phenotype. Cancer Cell. 2003;4:121–31.CrossRefPubMedGoogle Scholar
  54. 54.
    Van der Auwera I, Elst HJ, Van Laere SJ, et al. The presence of circulating total DNA and methylated genes is associated with circulating tumour cells in blood from breast cancer patients. Br J Cancer. 2009;100(8):1277–86.CrossRefPubMedGoogle Scholar
  55. 55.
    Hawkins N, Norrie M, Cheong K, et al. CpG island methylation in sporadic colorectal cancers and its relationship to microsatellite instability. Gastroenterology. 2002;122:1376–87.CrossRefPubMedGoogle Scholar
  56. 56.
    Samowitz WS, Albertsen H, Herrick J, et al. Evaluation of a large, population-based sample supports a CpG island methylator phenotype in colon cancer. Gastroenterology. 2005;129:837–45.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2010

Authors and Affiliations

  • Feng Jing
    • 1
    • 2
  • Wang Yuping
    • 2
  • Chen Yong
    • 3
  • Luo Jie
    • 3
  • Lu Jun
    • 3
  • Tang Xuanbing
    • 3
  • Hu Lihua
    • 1
  1. 1.Department of Laboratory Medicine, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Southern Medical UniversityFengxian HospitalShanghaiChina
  3. 3.Taihe HospitalYunyang Medical CollegeShiyanChina

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