Biomedical papers, 2013 (vol. 157), issue 4

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2013, 157(4):298-303 | 10.5507/bp.2013.035

Methylation analysis of tumor suppressor genes in endometroid carcinoma of endometrium using MS-MLPA

Eva Dvorakovaa, Marcela Chmelarovab, Jan Lacoc, Vladimir Palickab, Jiri Spaceka
a Department of Obstetrics and Gynecology, Faculty of Medicine in Hradec Kralove, Charles University in Prague and University Hospital Hradec Kralove, Hradec Kralove, Czech Republic
b Institute for Clinical Biochemistry and Diagnostics, Faculty of Medicine in Hradec Kralove, Charles University in Prague and University Hospital Hradec Kralove, Hradec Kralove
c The Fingerland Department of Pathology, Faculty of Medicine in Hradec Kralove, Charles University in Prague and University Hospital Hradec Kralove, Hradec Kralove

Background: Epigenetic changes are considered to be a frequent event during tumor development. Hypermethylation of promoter CpG islands represents an alternative mechanism for inactivation of tumor suppressor genes, DNA repair genes, cell cycle regulators and transcription factors. The aim of this study was to investigate promoter methylation of specific genes in endometrial cancer by comparison with normal endometrial tissue.

Materials and Methods: We used MS-MLPA (Methylation-specific Multiplex ligation-dependent probe amplification) to compare the methylation status of 59 tissue samples of endometroid type of endometrial carcinoma with 20 control samples of non-neoplastic endometrium.

Results: Using 15% cut-off for methylation, we observed significantly higher methylation in the CDH13 gene in endometrial cancer group. We observed significantly higher methylation in both WT1 and GATA5 genes in IB stage of endometroid carcinoma. We also observed significantly higher methylation in GATA5 gene in the group of poorly differentiated endometroid carcinoma.

Conclusion: The findings suggest the importance of hypermethylation of CDH13, WT1 and GATA5 genes in endometrial carcinogenesis and could have implications for future diagnostic and therapeutic strategies of endometrial cancer based on epigenetic changes.

Keywords: MS-MLPA, DNA methylation, endometrial cancer, CDH13, WT1, GATA5, epigenetics

Received: November 28, 2012; Accepted: April 26, 2013; Prepublished online: May 29, 2013; Published: December 12, 2013


References

  1. Liu F-S. Molecular carcinogenesis of endometrial cancer. Taiwaneses J Obstet Gynecol 2007;46:26-32. Go to original source...
  2. Sherman ME, Bur ME, Kurman RJ. p53 in endometrial cancer and its putative precursors: evidence for diverse pathways of tumorogenesis. Hum Pathol 1995;26:1268-74. Go to original source... Go to PubMed...
  3. Jones PA, Baylin SB. The epigenomics of cancer. Cell 2007;128:683-92. Go to original source... Go to PubMed...
  4. Clark SJ, Melki J. DNA methylation and gene silencing in cancer: which is the guilty party? Oncogene 2002;21:5380-7. Go to original source... Go to PubMed...
  5. Whitcomb BP, Mutch DG, Herzog TJ, Rader JS, Gibb RK, Goodfellow PJ. Frequent HOXA11 and THBS2 promoter methylation, and a methylator phenotype in endometrial adenocarcinoma. Clin Cancer Res 2003;9:2277-87. Go to PubMed...
  6. Mhawech P, Benz A, Cerato C, Greloz V, Assaly M, Desmond JC, Koeffler HP, Lodygin D, Hermeking H, Herrmann F, Schwaller J. Downregulation of 14-3-3sigma in ovary, prostate and endometrial carcinomas is associated with CpG island methylation. Mod Pathol 2005;18:340-8. Go to original source... Go to PubMed...
  7. Yeh KT, Yang MY, Liu TC, Chen JC, Chan WL, Lin SF, Chang JG. Abnormal expression of period 1 (PER1) in endometrial carcinoma. J Pathol 2005;206:111-20. Go to original source... Go to PubMed...
  8. Yoshida H, Broaddus R, Cheng W, Xie S, Naora H. Deregulation of the HOXA10 homeobox gene in endometrial carcinoma: role in epithelial-mesenchymal transition. Cancer Res 2006;66:889-97. Go to original source... Go to PubMed...
  9. Chmelarova M, Palicka V. The most frequent methods used for DNA methylation analysis. Cas Lek Cesk 2011;150:442-5. Go to PubMed...
  10. Nygren AO, Ameziane N, Duarte HM, Vijzelaar RN, Waisfisz Q, Hess CJ, Schouten JP, Errami A. 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. Go to original source... Go to PubMed...
  11. Tavassoli F.A, Devilee P. (Eds.): (2003) World Health Organization Classification of Tumors. Pathology and Genetics of Tumors of the Breast and Female Genital Organs. IARC Press: Lyon 2003:113-202.
  12. Moelans CB, Verschuur-Maes AH, van Diest PJ. Frequent promoter hypermethylation of BRCA2, CDH13, MSH6, PAX5, PAX6 and WT1 in ductal carcinoma in situ and invasive breast cancer. J Pathol 2011;225:222-31. Go to original source... Go to PubMed...
  13. Bol GM, Suijkerbuijk KP, Bart J, Vooijs M, van der Wall E, van Diest PJ. Methylation profiles of hereditary and sporadic ovarian cancer. Histopathology 2010;57:363-70. Go to original source... Go to PubMed...
  14. Seeber LM, Zweemer RP, Marchionni L, Massuger LF, Smit VT, van Baal WM, Verheijen RH, van Diest PJ. Methylation profiles of endometrioid and serous endometrial cancers. Endocr Relat Cancer 2010;17:663-73. Go to original source... Go to PubMed...
  15. Di Domenico M, Santoro A, Ricciardi C, Iaccarino M, Iaccarino S, Freda M, Feola A, Sanguedolce F, Losito S, Pasquali D, Di Spiezio Sardo A, Bifulco G, Nappi C, Bufo P, Guida M, De Rosa G, Abbruzzese A, Caraglia M, Pannone G. Epigenetic fingerprint in endometrial carcinogenesis: the hypothesis of a uterine field cancerization. Cancer Biol Ther 2011;12:447-57. Go to original source... Go to PubMed...
  16. Saito T, Nishimura M, Yamasaki H, Kudo R. Hypermethylation in promoter region of E-cadherin gene is associated with tumor dedifferention and myometrial invasion in endometrial carcinoma. Cancer 2003;97:1002-9. Go to original source... Go to PubMed...
  17. Banno K, Yanokura M, Susumu N, Kawaguchi M, Hirao N, Hirasawa A, Tsukazaki K, Aoki D. Relationship of the aberrant DNA hypermethylation of cancer-related genes with carcinogenesis of endometrial cancer. Oncol Rep 2006;16:1189-96. Go to original source... Go to PubMed...
  18. Pijnenborg JM, Kisters N, van Engeland M, Dunselman GA, de Haan J, de Goeij AF, Groothuis PG. APC, beta-catenin and E-cadherin and the development of recurrent endometrial carcinoma. Int J Gynecol Cancer 2004;14:947-56. Go to original source... Go to PubMed...
  19. Sneath RJ, Mangham DC. The normal structure and function of CD44 and its role in neoplasia. Mol Pathol 1998;51:191-200. Go to original source... Go to PubMed...
  20. Leblanc M, Poncelet C, Soriano D, Walker-Combrouze F, Madelenat P, Scoazec JY, Darai E. Alteration of CD44 and cadherins expression: possible association with augmented aggressiveness and invasiveness of endometrial carcinoma. Virchows Arch 2001;438:78-85. Go to original source... Go to PubMed...
  21. Ayhan A, Tok EC, Bildirici I, Ayhan A. Overexpression of CD44 variant 6 in human endometrial cancer and its prognostic significance. Gynecol Oncol 2001;80:355-8. Go to original source... Go to PubMed...
  22. Stokes GN, Shelton JB Jr, Zahn CM, Kendall BS. Association of CD44 isoform immunohistochemical expression with myometrial and vascular invasion in endometrioid endometrial carcinoma. Gynecol Oncol 2002;84:558-61. Go to original source... Go to PubMed...
  23. Afify AM, Craig S, Paulino AF, Stern R. Expression of hyaluronic acid and its receptors, CD44s and CD44v6, in normal, hyperplastic, and neoplastic endometrium. Ann Diagn Pathol 2005;9:312-8. Go to original source... Go to PubMed...
  24. Hong SC, Song JY, Lee JK, Lee NW, Kim SH, Yeom BW, Lee KW. Significance of CD44v6 expression in gynecologic malignancies. J Obstet Gynaecol Res 2006;32:379-86. Go to original source... Go to PubMed...
  25. Saegusa M, Hashimura M, Okayasu I. CD44 expression in normal, hyperplastic, and malignant endometrium. J Pathol 1998;184:297-306. Go to original source... Go to PubMed...
  26. Dohi S, Ohno S, Ohno Y, Kyo S, Soma G, Sugiyama H, Inoue M. WT1 expression correlates with angiogenesis in endometrial cancer tissue. Anticancer Res 2010;30:3187-92. Go to PubMed...
  27. Ohno S, Dohi S, Ohno Y, Kyo S, Sugiyama H, Suzuki N, Inoue M. Immunohistochemical detection of WT1 protein in endometrial cancer. Anticancer Res 2009;29:1691-5. Go to PubMed...
  28. Patient RK, McGhee JD. The GATA family (vertebrates and invertebrates). Curr Opin Genet Dev 2002;12:416-22. Go to original source... Go to PubMed...
  29. Molkentin JD. The zinc finger‑containing transcription factors GATA-4, -5, and -6: Ubiquitously expressed regulators of tissue‑specific gene expression. J Biol Chem 2000;275:38949-52. Go to original source... Go to PubMed...
  30. Guo M, Akiyama Y, House MG, Hooker CM, Heath E, Gabrielson E, Yang SC, Han Y, Baylin SB, Herman JG, Brock MV. Hypermethylation of the GATA genes in lung cancer. Clin Cancer Res 2004;10:7917-24. Go to original source... Go to PubMed...
  31. Guo M, House MG, Akiyama Y, Qi Y, Capagna D, Harmon J, Baylin SB, Brock MV, Herman JG. Hypermethylation of the GATA gene family in esophageal cancer. Int J Cancer 2006;119:2078-83. Go to original source... Go to PubMed...
  32. Akiyama Y, Watkins N, Suzuki H, Jair KW, van Engeland M, Esteller M, Sakai H, Ren CY, Yuasa Y, Herman JG, Baylin SB. GATA-4 and GATA-5 transcription factor genes and potential downstream antitumor target genes are epigenetically silenced in colorectal and gastrin cancer. Mol Cell Biol 2003;23:8429-39. Go to original source... Go to PubMed...
  33. Wakana K, Akiyama Y, Aso T, Yuasa Y. Involvement of GATA‑4/‑5 transcription factors in ovarian carcinogenesis. Cancer Lett 2006;241:281-8. Go to original source... Go to PubMed...