Detection of OPCML methylation, a possible epigenetic marker, from free serum circulating DNA to improve the diagnosis of early-stage ovarian epithelial cancer

Wang,Bi; Yu,Lei; Luo,Xin; Huang,Lin; Li,Qin‑Shan; Shao,Xiao‑Shan; Liu,Yi; Fan,Yu; Yang,Guo‑Zhen

doi:10.3892/ol.2017.6111

Detection of OPCML methylation, a possible epigenetic marker, from free serum circulating DNA to improve the diagnosis of early-stage ovarian epithelial cancer

Authors:
- Bi Wang
- Lei Yu
- Xin Luo
- Lin Huang
- Qin‑Shan Li
- Xiao‑Shan Shao
- Yi Liu
- Yu Fan
- Guo‑Zhen Yang
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Affiliations: School of Clinical Laboratory Science, Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China, Department of Obstetrics and Gynecology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China, Prenatal Diagnostic Center, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China, Department of Obstetrics and Gynecology, Guiyang Maternal and Child Health‑Care Hospital, Guiyang, Guizhou 550003, P.R. China
Published online on: May 3, 2017 https://doi.org/10.3892/ol.2017.6111
Pages: 217-223
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to identify the appropriate DNA sequence and design high‑quality primers for methylation-specific polymerase chain reaction (MSP). These primers may be used to examine and identify patients with early‑stage epithelial ovarian carcinoma (EOC). Opioid binding protein/cell adhesion molecule like (OPCML), Runt‑related transcription factor 3 and tissue factor pathway inhibitor 2 were selected as possible molecular markers. MSP primer sets were designed to monitor the methylation of the three markers. Free circulating DNA (fcDNA) from 194 patients with epithelial ovarian carcinoma and healthy donors were templates in the nested MSP. OPCML MSP was effective with respect to screening methylated fcDNA. One‑way ANOVA P‑values indicated that the difference in cancer antigen 125 (CA125), a biomarker for EOC diagnosis, level between early EOC and healthy donors was not significant. The methylation of OPCML was significantly altered in early‑stage EOC compared with healthy donors (P<0.0001), and this supported the hypothesis that specific fcDNA methylation was able to distinguish patients with early‑stage EOC from healthy donors. With respect to detecting early EOC, compared with the results of the CA125 test, MSP increased the κ coefficient from 0.140 to 0.757. Therefore, OPCML combined with fcDNA may be used to establish an improved clinical assay compared with the current CA125 test.

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1	American Cancer Society, . Cancer Facts & Figures 2015. American Cancer Society; Atlanta, GA: 2015
2	Ries LAG, Young JL, Keel GE, Eisner MP, Lin YD and Horner MJ: SEER Survival Monograph: Cancer Survival Among Adults. U.S. Seer Program, 1988–2001, Patient And Tumor Characteristics. National Cancer Institute, Seer Program, NIH Pub. No. 07-6215; Bethesda, MD: 2007
3	Prat J; FIGO Committee on Gynecologic Oncology, : Staging classification for cancer of the ovary, fallopian tube and peritoneum. Int J Gynaecol Obstet. 124:1–5. 2014. View Article : Google Scholar : PubMed/NCBI
4	Seiden MV: Gynecologic malignanciesHarrison's Principles of Internal Medicine. Longo DL, Kasper DL, Jameson JL, Fauci AS, Hauser SL and Loscalzo J: 1. 18th. McGraw-Hill; New York, NY: pp. 810–816. 2012
5	Suh KS, Park SW, Castro A, Patel H, Blake P, Liang M and Goy A: Ovarian cancer biomarkers for molecular biosensors and translational medicine. Expert Rev Mol Diagn. 10:1069–1083. 2010. View Article : Google Scholar : PubMed/NCBI
6	Gupta D and Lis CG: Role of CA125 in predicting ovarian cancer survival-a review of the epidemiological literature. J Ovarian Res. 2:132009. View Article : Google Scholar : PubMed/NCBI
7	Ozols RF: The current role of gemcitabine in ovarian cancer. Semin Oncol. 28 2 Suppl 7:S18–S24. 2001. View Article : Google Scholar
8	Leon SA, Shapiro B, Sklaroff DM and Yaros MJ: Free DNA in the serum of cancer patients and the effect of therapy. Cancer Res. 37:646–650. 1977.PubMed/NCBI
9	Flamini E, Mercatali L, Nanni O, Calistri D, Nunziatini R, Zoli W, Rosetti P, Gardini N, Lattuneddu A, Verdecchia GM and Amadori D: Free DNA and carcinoembryonic antigen serum levels: An important combination for diagnosis of colorectal cancer. Clin Cancer Res. 12:6985–6988. 2006. View Article : Google Scholar : PubMed/NCBI
10	Silva JM, Dominguez G, Garcia JM, Gonzalez R, Villanueva MJ, Navarro F, Provencio M, San Martin S, España P and Bonilla F: Presence of tumor DNA in plasma of breast cancer patients: Clinicopathological correlations. Cancer Res. 59:3251–3256. 1999.PubMed/NCBI
11	Gordian E, Ramachandran K, Reis IM, Manoharan M, Soloway MS and Singal R: Serum free circulating DNA is a useful biomarker to distinguish benign versus malignant prostate disease. Cancer Epidemiol Biomarkers Prev. 19:1984–1991. 2010. View Article : Google Scholar : PubMed/NCBI
12	Kamat AA, Bischoff FZ, Dang D, Baldwin MF, Han LY, Lin YG, Merritt WM, Landen CN Jr, Lu C, Gershenson DM, et al: Circulating cell-free DNA: A novel biomarker for response to therapy in ovarian carcinoma. Cancer Biol Ther. 5:1369–1374. 2006. View Article : Google Scholar : PubMed/NCBI
13	Mazurek AM, Fiszer-Kierzkowska A, Rutkowski T, Składowski K, Pierzyna M, Scieglińska D, Woźniak G, Głowacki G, Kawczyński R and Małusecka E: Optimization of circulating cell-free DNA recovery for KRAS mutation and HPV detection in plasma. Cancer Biomark. 13:385–394. 2013. View Article : Google Scholar : PubMed/NCBI
14	Ehrlich M, Gama-Sosa MA, Huang LH, Midgett RM, Kuo KC, McCune RA and Gehrke C: Amount and distribution of 5-methylcytosine in human DNA from different types of tissues of cells. Nucleic Acids Res. 10:2709–2721. 1982. View Article : Google Scholar : PubMed/NCBI
15	Jabbari K and Bernardi G: Cytosine methylation and CpG, TpG (CpA) and TpA frequencies. Gene. 333:143–149. 2004. View Article : Google Scholar : PubMed/NCBI
16	Deaton AM and Bird A: CpG islands and the regulation of transcription. Genes Dev. 25:1010–1022. 2011. View Article : Google Scholar : PubMed/NCBI
17	Kim TY, Lee HJ, Hwang KS, Lee M, Kim JW, Bang YJ and Kang GH: Methylation of RUNX3 in various types of human cancers and premalignant stages of gastric carcinoma. Lab Invest. 84:479–484. 2004. View Article : Google Scholar : PubMed/NCBI
18	Zhang S, Wei L, Zhang A, Zhang L and Yu H: RUNX3 gene methylation in epithelial ovarian cancer tissues and ovarian cancer cell lines. OMICS. 13:307–311. 2009. View Article : Google Scholar : PubMed/NCBI
19	Sierko E, Wojtukiewicz MZ and Kisiel W: The role of tissue factor pathway inhibitor-2 in cancer biology. Semin Thromb Hemost. 33:653–659. 2007. View Article : Google Scholar : PubMed/NCBI
20	Sellar GC, Watt KP, Rabiasz GJ, Stronach EA, Li L, Miller EP, Massie CE, Miller J, Contreras-Moreira B, Scott D, et al: OPCML at 11q25 is epigenetically inactivated and has tumor-suppressor function in epithelial ovarian cancer. Nat Genet. 34:337–343. 2003. View Article : Google Scholar : PubMed/NCBI
21	Chuang LS and Ito Y: RUNX3 is multifunctional in carcinogenesis of multiple solid tumors. Oncogene. 29:2605–2615. 2010. View Article : Google Scholar : PubMed/NCBI
22	Herman JG, Graff JR, Myöhänen S, Nelkin BD and Baylin SB: Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA. 93:pp. 9821–9826. 1996; View Article : Google Scholar : PubMed/NCBI
23	Li LC and Dahiya R: MethPrimer: Designing primers for methylation PCRs. Bioinformatics. 18:1427–1431. 2002. View Article : Google Scholar : PubMed/NCBI
24	Ledermann JA, Raja FA, Fotopoulou C, Gonzalez-Martin A, Colombo N and Sessa C; ESMO Guidelines Working Group, : Newly diagnosed and relapsed epithelial ovarian carcinoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol. 24 Suppl 6:vi24–vi32. 2013. View Article : Google Scholar : PubMed/NCBI
25	Sung CK, Li D, Andrews E, Drapkin R and Benjamin T: Promoter methylation of the SALL2 tumor suppressor gene in ovarian cancers. Mol Oncol. 7:419–427. 2013. View Article : Google Scholar : PubMed/NCBI
26	Katsaros D, Cho W, Singal R, Fracchioli S, De La Longrais IA Rigault, Arisio R, Massobrio M, Smith M, Zheng W, Glass J and Yu H: Methylation of tumor suppressor gene p16 and prognosis of epithelial ovarian cancer. Gynecol Oncol. 94:685–692. 2004. View Article : Google Scholar : PubMed/NCBI
27	Ozdemir F, Altinisik J, Karateke A, Coksuer H and Buyru N: Methylation of tumor suppressor genes in ovarian cancer. Exp Ther Med. 4:1092–1096. 2012.PubMed/NCBI