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Application of Multiplex Nested Methylated Specific PCR in Early Diagnosis of Epithelial Ovarian Cancer

  • Wang, Bi (Guiyang Medical University) ;
  • Yu, Lei (The First School of Medicine, The First Affiliated Hospital of Jinan University) ;
  • Yang, Guo-Zhen (Guiyang Medical University) ;
  • Luo, Xin (Department of Obstetrics and Gynecology, The First Affiliated Hospital of Jinan University) ;
  • Huang, Lin (Department of Obstetrics and Gynecology, Affiliated Hospital of Guiyang Medical University)
  • Published : 2015.04.14

Abstract

Objective: To explore the application of multiplex nested methylated specific polymerase chain reaction (PCR) in the early diagnosis of epithelial ovarian carcinoma (EOC). Materials and Methods: Serum and fresh tissue samples were collected from 114 EOC patients. RUNX3, TFPI2 and OPCML served as target genes. Methylation levels of tissues were assessed by multiplex nested methylated specific PCR, the results being compared with those for carcinoma antigen 125 (CA125). Results: The serum free deoxyribose nucleic acid (DNA) methylation spectrum of EOC patients was completely contained in the DNA spectrum of cancer tissues, providing an accurate reflection of tumor DNA methylation conditions. Serum levels of CA125 and free DNA methylation in the EOC group were evidently higher than those in benign lesion and control groups (p<0.05). Patients with early EOC had markedly lower serum CA125 than those with advanced EOC (p<0.05), but there was no significant difference in free DNA methylation (p>0.05). The sensitivity, specificity and positive predicative value (PPV) of multiplex nested methylated specific PCR were significantly higher for detection of all patients and those with early EOC than those for CA125 (p<0.05). In the detection of patients with advanced EOC, the PPV of CA125 detection was obviously lower than that of multiplex nested methylated specific PCR (p>0.05), but there was no significant difference in sensitivity (p>0.05). Conclusions: Serum free DNA methylation can be used as a biological marker for EOC and multiplex nested methylated specific PCR should be considered for early diagnosis since it can accurately determine tumor methylation conditions.

Keywords

DNA methylation;multiplex PCR;nested PCR;epithelial ovarian cancer;early diagnosis

Acknowledgement

Supported by : Academician Workstation Construction Projects of Guangdong Province, International Cooperation Projects of Guangdong Province

References

  1. Amornpisutt R, Proungvitaya S, Jearanaikoon P, et al (2015). DNA methylation level of OPCML and SFRP1: a potential diagnostic biomarker of cholangiocarcinoma. Tumour Biol, (in press).
  2. Buys SS, Partridge E, Black A, et al (2011). Effect of screening on ovarian cancer mortality: the prostate, lung, colorectal and ovarian (PLCO) cancer screening randomized controlled trial. JAMA, 305, 2295-303. https://doi.org/10.1001/jama.2011.766
  3. Clarke-Pearson DL (2009). Screening for ovarian cancer. New Engl J Med, 361, 170-7. https://doi.org/10.1056/NEJMcp0901926
  4. Chen H, Zhang C, Sheng Y, et al (2014). Frequent SOCS3 and 3OST2 promoter methylation and their epigenetic regulation in endometrial carcinoma. Am J Cancer Res, 5, 180-90.
  5. Guo S, Yan F, Xu J, et al (2015). Identification and validation of the methylation biomarkers of non-small cell lung cancer (NSCLC). Clin Epigenetics, 7, 3. https://doi.org/10.1186/s13148-014-0035-3
  6. Ji T, Zheng ZG, Wang FM, et al (2014). Differential microRNA expression by Solexa sequencing in the sera of ovarian cancer patients. Asian Pac J Cancer Prev, 15, 1739-43. https://doi.org/10.7314/APJCP.2014.15.4.1739
  7. Karami S, Andreotti G, Liao LM, et al (2015). LINE-1 methylation levels in pre-diagnostic leukocyte DNA and future renal cell carcinoma risk. Epigenetics, (in press).
  8. Liggett TE, Melnikov A, Yi Q, et al (2011). Distinctive DNA methylation patterns of cell-free plasma DNA in women with malignant ovarian tumors. Gynecol Oncol, 120, 113-20. https://doi.org/10.1016/j.ygyno.2010.09.019
  9. McIntosh MW, Liu Y, Drescher C, et al (2007). Validation and characterization of human kallikrein 11 as a serum marker for diagnosis of ovarian carcinoma. Clin Cancer Res, 13, 4422-8. https://doi.org/10.1158/1078-0432.CCR-06-2224
  10. Nelissen EC, van Montfoort AP, Dumoulin JC, et al (2011). Epigenetics and the placenta. Hum Reprod Update, 17, 397-417. https://doi.org/10.1093/humupd/dmq052
  11. Nordlund J, Bäcklin CL, Zachariadis V, et al (2015). DNA methylation-based subtype prediction for pediatric acute lymphoblastic leukemia. Clin Epigenetics, 7, 11. https://doi.org/10.1186/s13148-014-0039-z
  12. Pothuri B, Leitao MM, Levine DA, et al (2010). Genetic analysis of the early natural history of epithelial ovarian carcinoma. PLoS One, 5, 10358. https://doi.org/10.1371/journal.pone.0010358
  13. Shih IeM, Kurman RJ (2004). Ovarian tumorigenesis: a proposed model based on morphological and molecular genetic analysis. Am J Pathol, 164, 1511-8. https://doi.org/10.1016/S0002-9440(10)63708-X
  14. Siegel R, Deepa Naishadham MA, Ahmedin Jemal DVM (2013). Cancer statistics, 2013. CA Cancer J Clin, 63, 11-30. https://doi.org/10.3322/caac.21166
  15. Tost J (2010). DNA methylation: an introduction to the biology and the disease-associated changes of a promising biomarker. Mol Biotechnol, 44, 71-81. https://doi.org/10.1007/s12033-009-9216-2
  16. The Department of Health, Australian Government (2011). Cancer facts and figures. http://www.health.gov.au/internet/main/publishing.nsf/Content/cancer-facts
  17. Tiwawech D, Srisuttee R, Rattanatanyong P, et al (2014). Alu methylation in serum from patients with nasopharyngeal carcinoma. Asian Pac J Cancer Prev, 15, 9797-800. https://doi.org/10.7314/APJCP.2014.15.22.9797
  18. Wang B, Liu SZ, Zheng RS, et al (2014).Time trends of ovarian cancer incidence in China. Asian Pac J Cancer Prev, 15, 191-3. https://doi.org/10.7314/APJCP.2014.15.1.191
  19. Yu Shaorong, Liu Baorui, Lu Jianwei, et al (2013). Circulating DNA as potential biomarker for cancer individualized therapy. J Int Transl Med, 1, 144-51.
  20. Yamashita K, Waraya M, Kim MS, et al (2014). Detection of methylated CDO1 in plasma of colorectal cancer; a PCR study. PLoS One, 9, 113546. https://doi.org/10.1371/journal.pone.0113546
  21. Zhou F, Ma M, Tao G, et al (2014). Detection of circulating methylated opioid binding protein/cell adhesion moleculelike gene as a biomarker for ovarian carcinoma. Clin Lab, 60, 759-65.