Methylation of selected tumor-supressor genes in benign and malignant ovarian tumors

Authors: M. Cuľbová 1;  Z. Lasabová 2;  A. Štanclová 2;  P. Tilandyová 2;  P. Žúbor 1;  R. Fiolka 1;  J. Danko 1;  J. Višňovský 1
Authors‘ workplace: Gynekologicko-pôrodnícka klinika JLF UK a UNM, Martin, prednosta prof. MUDr. J. Danko, CSc. 1;  Ústav molekulovej biológie, Vrútky, prednostka RNDr. Z. Lasabová, Ph. D. 2
Published in: Ceska Gynekol 2011; 76(4): 274-279


To evaluate the usefullness of examination of methylation status of selected tumor-supressor genes in early diagnosis of ovarian cancer.

Prospective clinical study.

Department of Gynecology and Obstetrics, Department of Molecular Biology, Jessenius Medical Faculty, Commenius University, Martin, Slovak Republic.

In this study we analyzed hypermethylation of 5 genes RASSF1A, GSTP, E-cadherin, p16 and APC in ovarian tumor samples from 34 patients – 13 patients with epithelial ovarian cancer, 2 patients with border-line ovarian tumors, 12 patients with benign lesions of ovaries and 7 patients with healthy ovarian tissue. The methylation status of promoter region of tumor-supressor genes was determined by Methylation Specific Polymerase Chain Reaction (MSP) using a nested two-step approach with bisulfite modified DNA template and specific primers.

Gene methylation analysis revealed hypermethylation of gene RASSF1A (46%) and GSTP (8%) only in malignant ovarian tissue samples. Ecad, p16 and APC genes were methylated both in maignant and benign tissue samples. Methylation positivity in observed genes was present independently to all clinical stages of ovarian cancer and to tumor grades. However, there was observed a trend of increased number and selective involvement of methylated genes with increasing disease stages. Furthermore, there was no association between positive methylation status and histological subtypes of ovarian carcinomas.

RASSF1A and GSTP promoter methylation positivity is associated with ovarian cancer. The revealed gene-selective methylation positivity and the increased number of methylated genes with advancing disease stages could be considered as a useful molecular marker for early detection of ovarian cancer. However, there is need to find diagnostic approach of specifically and frequently methylated genes to determining a methylation phenotype for early detection of ovarian malignancies.

Key words:
ovarian cancer, DNA methylation, early detection.


1. Hoskins, WJ., Perez, CA., Young, RC., et al. Principles and practice of gynecologic oncology, 4th ed. Philadelphia: Lippincott Williams and Wilkins, 2005, p. 9-13, 33-36, 109-113, 157-167, 190-193, 250-259, 895-987.

2. Woodward, ER., Sleightholme, HV., Considine, AM., et al. Annual surveillance by CA 125 and transvaginal ultrasound for ovarian cancer in both high-risk and population risk women is ineffective. BJOG, 2007, 114, p. 1500-1509.

3. Das, PM., Singal, R. DNA methylation and cancer. J Clin. Oncol, 2004, 22, p. 4632-4642.

4. Paluszczak, J., Baer-Dubowska, W. Epigenetic diagnostics of cancer – the application of DNA methylation markers. J Appl Genet, 2006, 47, p. 365-375.

5. Wiley, A., Katsaros, D., Chen, H., et al. Abberant promoter methylation of multiple genes in malignant ovarian tumors and in ovarian tumors with low malignant potential. Cancer, 2006, 107, p. 299-308.

6. Watts , GS., Futscher, BW., Holtan, N., et al. DNA methylation changes in ovarian cancer are cumulative with disease progression and identify tumor stage. BMC Med Genomics, 2008, 1, p. 47-59.

7. Barton, CA., Hacker, NF., Clark, SJ., et al. DNA methylation changes in ovarian cancer: implications for early diagnosis, prognosis and treatment. Gynec Oncol, 2008, 109, p. 129-139.

8. Vos, MD., Martinez, A., Elam, C., et al. A role for the RASSF1A tumor supressor in the regulation of tubulin polymerization and genomic stability. Cancer Res, 2004, 64, p. 4244-4250.

9. Van der Weyden, L., Adams, DJ. The Ras-association domain family (RASSF) members and their role in human tumorigenesis. Biochim Biophys Acta, 2007, 1776, p. 58-85.

10. Xiao, L., Yu, L., Can, M. Expression of beta-catenin and APC protein in ovarian epithelial tumor and its implication. Chinese J Cancer Res, 2007, 19, p. 72-75.

11. Su, HY., Lai, HC., Lin, YW., et al. An epigenetic marker panel for screening and prognostic prediction of ovarian cancer. Int J Cancer, 2009, 124, p. 387-393.

12. Pfeifer, GP., Dammann, R. Methylation of the tumor supressor gene RASSF1A in human tumors. Biochemistry, 2005, 70, p. 699-707.

13. Ma, L., Guo, Q., Ma, Y., et al. Clinicopathological implications of inactivation of RASSF1A in serous epithelial ovarian cancers. Eur J Gynaecol Oncol, 2009, 30, p. 370-374.

14. Ma, L., Zhang, JH., Liu, FR., et al. Hypermethylation of promoter region of RASSF1A gene in ovarian malignant epithelial tumors. Zhongua Zhong Liu Za Zhi, 2005, 27, p. 657‑659.

15. Choi, YL., Kang, SY., Shin, YK., et al. Abberant hypermethylation of RASSF1A promoter in ovarian borderline tumors and carcinomas. Virchows Arch, 2006, 448, p. 331-336.

16. Yuecheng, Y., Hongmei, L., Xiaoyan, X. Clinical evaluation of E-cadherin expression and its regulation mechanism in epithelial ovarian cancer. Clin Exp Metastasis, 2006, 23, p. 65-74.

17. Koensgen, D., Freitag, C., Klaman, I., et al. Expression and localization of e-cadherin in epithelial ovarian cancer. Anticancer Res, 2010, 30, p. 2525-2530.

18. Liggett, TE., Melnikov, A., Yi, Q., et al. Distinctive DNA methylation patterns of cell-free plasma DNA in women, with malignant ovarian tumors. Gynecol Oncol, 2011, 120, p. 113-120.

19. Houshdara, S., Hawley, S., Palmer, C., et al. DNA methylation profiles of ovarian epithelial carcinoma tumors and cell lines. PLoS ONE, 2010, 5, e9359.

20. Balch, C., Fang, F., Matei, DE., et al. Minireview: Epigenetic changes in ovarian cancer. Endocrinology, 2009, 150, p. 4003‑4011.

21. Strathdee, G. Epigenetic versus genetic alterations in the inactivation of E-cadherin. Cancer Biol, 2002, 12, p. 373-379.

22. Huang, ZW., Jansen, RA., Fabbry, E., et al. Identification of candidate epigenetic biomarkers for ovarian cancer detection. Oncol Res, 2009, 22, p. 853-861.

23. Teschendorff, AE., Menon, U., Gentry-Maharaj, A., et al. An epigenetic signature in peripheral blood predicts active ovarian cancer. PLoS ONE, 2009, 4, e8274.

24. Asadollahi, R., Hyde, CAC., Zhong, XY. Epigenetics of ovarian cancer: from lab to clinic. Gynecol Oncol, 2010, 118, p. 81-87.

25. Konstantinopoulos, PA., Spentzos, D., Cannistra, SA. Gene-expression profiling in epithelial ovarian cancer. Nature Clin Pract Oncol, 2008, 5, p. 577-587.

26. Helou, K., Padilla-Nash, H., Wangsa, D., et al. Comparative genome hybridization reveals specific genomic imbalances during the genesis from benign, through borderline to malignant ovarian tumors. Cancer Genet Cytogenet, 2006, 170, p. 1-8.

27. Makarla, PB., Soboorian, H., Ashfaq, R., et al. Promoter hypermethylation profile of ovarian epithelial neoplasms. Clin Cancer Res, 2005, 11, p. 5365-5369.

28. Melnikov, A., Scholtens, D., Godwin, A., et al. Differential methylation profile of ovarian cancer in tissues and plasma. J Molec Diagnost, 2009, 11, p. 60-65.

29. Bol, GB., Suijkerbuijk, PM., Bart, J., et al. Methylation profiles of hereditary and sporadic ovarian cancer. Histopathology, 2010, 57, p. 363-370.

30. Katsaros, D., Cho, W., Singal, R., et al. Methylation of tumor-supressor gene p16 and prognosis of epithelial ovarian cancer. Gynecol Oncol, 2004, 94, p. 685-692.

31. Mc Cluskey, LL., Chen, C., Delgadillo, E., et al. Differencies in p16 gene methylation and expression in benign and malignant ovarian tumors. Gynecol Oncol, 1999, 72, p. 87-92.

32. Auerkari, EI. Methylation of tumor-supressor genes p16 (INK4a), p27 (KIP1) and Ec-cadherin in carcinogenesis. Oral Oncol, 2006, 42, p. 5-13.

33. Dammann, R., Schagdarsurengin, U., Seidel, C., et al. The tumor supressor RASSF1A in human carcinogenesis: an update. Histol Histopathol, 2005, 20, p. 245-263.

34. House, MG., Guo, M., Iacobuzio-Donahue, C., et al. Molecular progression of promoter methylation in intraductal papillary mucosinous neoplasms (IPMN) of the pancreas. Carcinogenesis, 2003, 24, p. 193-198.

Paediatric gynaecology Gynaecology and obstetrics Reproduction medicine
Forgotten password

Enter the email address that you registered with. We will send you instructions on how to set a new password.


Don‘t have an account?  Create new account