Home SOX17 promoter methylation in plasma circulating tumor DNA of patients with non-small cell lung cancer
Article
Licensed
Unlicensed Requires Authentication

SOX17 promoter methylation in plasma circulating tumor DNA of patients with non-small cell lung cancer

  • Ioanna Balgkouranidou , Maria Chimonidou , Georgia Milaki , Emily Tsaroucha , Stylianos Kakolyris , Vasilis Georgoulias and Evi Lianidou EMAIL logo
Published/Copyright: January 7, 2016
Become an author with De Gruyter Brill
Submit Manuscript Author Information
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 54 Issue 8

Abstract

Background: SOX17 belongs to the high-mobility group-box transcription factor superfamily and down-regulates the Wnt pathway. The aim of our study was to evaluate the prognostic significance of SOX17 promoter methylation in circulating tumor DNA (ctDNA) in plasma of non-small cell lung cancer (NSCLC) patients.

Methods: We examined the methylation status of SOX17 promoter in 57 operable NSCLC primary tumors and paired adjacent non-cancerous tissues and in ctDNA isolated from 48 corresponding plasma samples as well as in plasma from 74 patients with advanced NSCLC and 49 healthy individuals. SOX17 promoter methylation was examined by Methylation Specific PCR (MSP).

Results: In operable NSCLC, SOX17 promoter was fully methylated in primary tumors (57/57, 100%), and in corresponding ctDNA (27/48, 56.2%) while it was detected in only 1/49 (2.0%) healthy individuals. In advanced NSCLC, SOX17 promoter was methylated in ctDNA in 27/74 (36.4%) patients and OS was significantly different in favor of patients with non-methylated SOX17 promoter (p=0.012). Multivariate analysis revealed that SOX17 promoter methylation in ctDNA was an independent prognostic factor associated with OS in patients with advanced but not operable NSCLC.

Conclusions: Our results show that SOX17 promoter is highly methylated in primary tumors and in corresponding plasma samples both in operable and advanced NSCLC. In the advanced setting, SOX17 promoter methylation in plasma ctDNA has a statistical significant influence on NSCLC patient’s survival time. Detection of SOX17 promoter methylation in plasma provides prognostic information and merits to be further evaluated as a circulating tumor biomarker in patients with operable and advanced NSCLC.

Keywords: cell free DNA; circulating tumor DNA; DNA methylation; methylation specific PCR; non-small cell lung cancer; prognostic significance; SOX17
Corresponding author: Prof. Evi Lianidou, Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, 15771, Greece, Phone: +30 210 7274319, Fax: +30 210 7274750, E-mail:

Acknowledgments

Ioanna Balgkouranidou is a recipient of a PhD studentship from the Hellenic Oncology Research Group (HORG). We would like to thank Dr. Dora Chatzidaki, for her significant help in the statistical analysis of our results. IB, MC, performed the experiments, GM, EGT, SK and VG provided the patients’ samples and all the clinical data needed, ES conceived and supervised the whole study and wrote the manuscript.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. Janssen-Heijnen ML, van Erning FN, De Ruysscher DK, Coebergh JW, Groen HJ. Variation in causes of death in patients with non-small cell lung cancer according to stage and time since diagnosis. Ann Oncol 2015;5:902–7.10.1093/annonc/mdv061Search in Google Scholar PubMed

2. Johnson DH, Schiller JH, Bunn PA Jr. Recent clinical advances in lung cancer management. J Clin Oncol 2014;32:973–82.10.1200/JCO.2013.53.1228Search in Google Scholar PubMed

3. Rami-Porta R, Bolejack V, Giroux DJ, Chansky K, Crowley J, Asamura H, et al. International Association for the Study of Lung Cancer Staging and Prognostic Factors Committee, Advisory Board Members and Participating Institutions. The IASLC lung cancer staging project: the new database to inform the eighth edition of the TNM classification of lung cancer. J Thorac Oncol 2014;9:1618–24.10.1097/JTO.0000000000000334Search in Google Scholar PubMed

4. Carter BW, Glisson BS, Truong MT, Erasmus JJ. Small cell lung carcinoma: staging, imaging, and treatment considerations. Radiographics 2014;34:1707–21.10.1148/rg.346140178Search in Google Scholar PubMed

5. Keedy VL, Temin S, Somerfield MR, Beasley MB, Johnson DH, McShane LM, et al. American Society of Clinical Oncology provisional clinical opinion: epidermal growth factor receptor (EGFR) Mutation testing for patients with advanced non-small-cell lung cancer considering first-line EGFR tyrosine kinase inhibitor therapy. J Clin Oncol 2011;29:2121–27.10.1200/JCO.2010.31.8923Search in Google Scholar PubMed

6. Stella GM, Scabini R, Inghilleri S, Cemmi F, Corso S, Pozzi E, et al. EGFR and KRAS mutational profiling in fresh non-small cell lung cancer (NSCLC) cells. J Cancer Res Clin Oncol 2013;139:1327–35.10.1007/s00432-013-1444-ySearch in Google Scholar PubMed

7. Balgkouranidou I, Liloglou T, Lianidou ES. Lung cancer epigenetics: emerging biomarkers. Biomark Med 2013;7:49–58.10.2217/bmm.12.111Search in Google Scholar PubMed

8. Lou-Qian Z, Rong Y, Ming L, Xin Y, Feng J, Lin X. The prognostic value of epigenetic silencing of p16 gene in NSCLC patients: a systematic review and meta-analysis. PLoS One 2013;8:e54970.10.1371/journal.pone.0054970Search in Google Scholar PubMed PubMed Central

9. Wang J, Lee JJ, Wang L, Liu DD, Lu C, Fan YH, et al. Value of p16INK4a and RASSF1A promoter hypermethylation in prognosis of patients with resectable non-small cell lung cancer. Clin Cancer Res 2004;10:6119–25.10.1158/1078-0432.CCR-04-0652Search in Google Scholar PubMed

10. Juergens RA, Wrangle J, Vendetti FP, Murphy SC, Zhao M, Coleman B, et al. Combination epigenetic therapy has efficacy in patients with refractory advanced non-small cell lung cancer. Cancer Discov 2011;1:598–607.10.1158/2159-8290.CD-11-0214Search in Google Scholar PubMed PubMed Central

11. Chiu RW, Lo YM. Clinical applications of maternal plasma fetal DNA analysis: translating the fruits of 15 years of research. Clin Chem Lab Med 2013;51:197–204.10.1515/cclm-2012-0601Search in Google Scholar PubMed

12. Marzese DM, Hirose H, Hoon DS. Diagnostic and prognostic value of circulating tumor-related DNA in cancer patients. Expert Rev Mol Diagn 2013;13:827–44.10.1586/14737159.2013.845088Search in Google Scholar PubMed

13. Aung KL, Board RE, Ellison G, Donald E, Ward T, Clack G, et al. Current status and future potential of somatic mutation testing from circulating free DNA in patients with solid tumours. Hugo J 2010;4:11–21.10.1007/s11568-011-9149-2Search in Google Scholar PubMed PubMed Central

14. Nie K, Jia Y, Zhang X. Cell-free circulating tumor DNA in plasma/serum of non-small cell lung cancer. Tumour Biol 2015;36:7–19.10.1007/s13277-014-2758-3Search in Google Scholar PubMed

15. Balgkouranidou I, Chimonidou M, Milaki G, Tsarouxa EG, Kakolyris S, Welch DR, et al. Breast cancer metastasis suppressor-1 promoter methylation in cell-free DNA provides prognostic information in non-small cell lung cancer. Br J Cancer 2014;110:2054–62.10.1038/bjc.2014.104Search in Google Scholar PubMed PubMed Central

16. Schwarzenbach H, Hoon DS, Pantel K. Cell-free nucleic acids as biomarkers in cancer patients. Nat Rev Cancer 2011;11:426–37.10.1038/nrc3066Search in Google Scholar PubMed

17. Fischer JR, Ohnmacht U, Rieger N, Zemaitis M, Stoffregen C, Manegold C, et al. Prognostic significance of RASSF1A promoter methylation on survival in non small cell lung cancer patients treated with gemcitabine. Lung Cancer 2007;56:115–23.10.1016/j.lungcan.2006.11.016Search in Google Scholar PubMed

18. Ponomaryova AA, Rykova EY, Cherdyntseva NV, Skvortsova TE, Dobrodeev AY, Zav’yalov AA, et al. Potentialities of aberrantly methylated circulating DNA for diagnostics and post-treatment follow-up of lung cancer patients. Lung Cancer 2013;81:397–403.10.1016/j.lungcan.2013.05.016Search in Google Scholar PubMed

19. Gubbay J, Collignon J, Koopman P, Capel B, Economou A, Münsterberg A, et al. A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature 1990;346:245–50.10.1038/346245a0Search in Google Scholar PubMed

20. Sinner D, Kordich JJ, Spence JR, Opoka R, Rankin S, Lin SC, et al. Sox17 and Sox4 differentially regulate beta-catenin/T-cell factor activity and proliferation of colon carcinoma cells. Mol Cell Biol 2007;27:7802–15.10.1128/MCB.02179-06Search in Google Scholar PubMed PubMed Central

21. Zhang W, Glöckner SC, Guo M, Machida EO, Wang DH, Easwaran H, et al. Epigenetic inactivation of the canonical Wnt antagonist SRY-box containing gene 17 in colorectal cancer. Cancer Res 2008;68:2764–72.10.1158/0008-5472.CAN-07-6349Search in Google Scholar PubMed PubMed Central

22. Fu DY, Wang ZM, Li-Chen, Wang BL, Shen ZZ, Huang W, et al. Sox17, the canonical Wnt antagonist, is epigenetically inactivated by promoter methylation in human breast cancer. Breast Cancer Res Treat 2010;119:601–12.10.1007/s10549-009-0339-8Search in Google Scholar PubMed

23. Chimonidou M, Strati A, Tzitzira A, Sotiropoulou G, Malamos N, Georgoulias V, et al. DNA methylation of tumor suppressor and metastasis suppressor genes in circulating tumor cells. Clin Chem 2011;57:1169–77.10.1373/clinchem.2011.165902Search in Google Scholar PubMed

24. Chimonidou M, Strati A, Malamos N, Georgoulias V, Lianidou ES. SOX17 promoter methylation in circulating tumor cells and matched cell-free DNA isolated from plasma of patients with breast cancer. Clin Chem 2013;59:270–9.10.1373/clinchem.2012.191551Search in Google Scholar PubMed

25. Kroupis C, Christopoulos K, Devetzoglou M, Tsiagas I, Lianidou ES. Asymmetric real-time PCR detection of BRCA1 mutation by melting curve analysis in the LightCycler. Clin Chim Acta 2008;390:141–4.10.1016/j.cca.2007.12.024Search in Google Scholar PubMed

26. Mastoraki S, Chimonidou M, Dimitrakopoulos L, Kounelis S, Malamos N, Georgoulias V, et al. A rapid and accurate closed-tube Methylation-Sensitive High Resolution Melting Analysis assay for the semi-quantitative determination of SOX17 promoter methylation in clinical samples. Clin Chim Acta 2015;444:303–9.10.1016/j.cca.2015.02.035Search in Google Scholar PubMed

27. Lianidou ES, Strati A, Markou A. Circulating tumor cells as promising novel biomarkers in solid cancers. Crit Rev Clin Lab Sci 2014;51:160–71.10.3109/10408363.2014.896316Search in Google Scholar PubMed

28. Faugeroux V, Pailler E, Auger N, Taylor M, Farace F. Clinical Utility of Circulating Tumor Cells in ALK-Positive Non-Small-Cell Lung Cancer. Front Oncol 2014;4:281.10.3389/fonc.2014.00281Search in Google Scholar PubMed PubMed Central

29. Marzese DM, Hoon DS. Emerging technologies for studying DNA methylation for the molecular diagnosis of cancer. Expert Rev Mol Diagn 2015;15:647–64.10.1586/14737159.2015.1027194Search in Google Scholar PubMed PubMed Central

30. Luke JJ, Oxnard GR, Paweletz CP, Camidge DR, Heymach JV, Solit DB, et al. Cell Free DNA Working Group. Realizing the potential of plasma genotyping in an age of genotype-directed therapies. J Natl Cancer Inst 2014;106:dju214.10.1093/jnci/dju214Search in Google Scholar PubMed PubMed Central

31. Madhavan D, Wallwiener M, Bents K, Zucknick M, Nees J, Schott S, et al. Plasma DNA integrity as a biomarker for primary and metastatic breast cancer and potential marker for early diagnosis. Breast Cancer Res Treat 2014;146:163–74.10.1007/s10549-014-2946-2Search in Google Scholar PubMed

32. Ilie M, Hofman V, Long E, Bordone O, Selva E, Washetine K, et al. Current challenges for detection of circulating tumor cells and cell-free circulating nucleic acids, and their characterization in non-small cell lung carcinoma patients. What is the best blood substrate for personalized medicine? Ann Transl Med 2014;2:107.Search in Google Scholar

33. Douillard JY, Ostoros G, Cobo M, Ciuleanu T, Cole R, McWalter G, et al. Gefitinib treatment in EGFR mutated caucasian NSCLC: circulating-free tumor DNA as a surrogate for determination of EGFR status. J Thorac Oncol 2014;9:1345–53.10.1097/JTO.0000000000000263Search in Google Scholar PubMed PubMed Central

34. Douillard JY, Ostoros G, Cobo M, Ciuleanu T, McCormack R, Webster A, et al. First-line gefitinib in Caucasian EGFR mutation-positive NSCLC patients: a phase-IV, open-label, single-arm study. Br J Cancer 2014;110:55–62.10.1038/bjc.2013.721Search in Google Scholar PubMed PubMed Central

35. Balgkouranidou I, Karayiannakis A, Matthaios D, Bolanaki H, Tripsianis G, Tentes AA, et al. Assessment of SOX17 DNA methylation in cell free DNA from patients with operable gastric cancer. Association with prognostic variables and survival. Clin Chem Lab Med 2013;51:1505–10.10.1515/cclm-2012-0320Search in Google Scholar PubMed

36. Du YC, Oshima H, Oguma K, Kitamura T, Itadani H, Fujimura T, et al. Induction and down-regulation of Sox17 and its possible roles during the course of gastrointestinal tumorigenesis. Gastroenterology 2009;137:1346–57.10.1053/j.gastro.2009.06.041Search in Google Scholar PubMed

37. Li Z, Guo X, Wu Y, Li S, Yan J, Peng L, et al. Methylation profiling of 48 candidate genes in tumor and matched normal tissues from breast cancer patients. Breast Cancer Res Treat 2015;149:767–79.10.1007/s10549-015-3276-8Search in Google Scholar PubMed

38. Stewart DJ. Wnt signaling pathway in non-small cell lung cancer. J Natl Cancer Inst 2014;106:djt356.10.1093/jnci/djt356Search in Google Scholar PubMed

Received: 2015-8-10
Accepted: 2015-11-1
Published Online: 2016-1-7
Published in Print: 2016-8-1

©2016 by De Gruyter

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. Laboratory analytical quality – the process continues
  4. Measurement uncertainty – a revised understanding of its calculation and use
  5. Review
  6. Substrate-zymography: a still worthwhile method for gelatinases analysis in biological samples
  7. Opinion Papers
  8. Is the combination of trueness and precision in one expression meaningful? On the use of total error and uncertainty in clinical chemistry
  9. The problem with total error models in establishing performance specifications and a simple remedy
  10. Measurement uncertainty for clinical laboratories – a revision of the concept
  11. Uncertainty in measurement and total error – are they so incompatible?
  12. General Clinical Chemistry and Laboratory Medicine
  13. Using the hazard ratio to evaluate allowable total error in predictive measurands
  14. Performance of electrolyte measurements assessed by a trueness verification program
  15. Analytical interference by monoclonal immunoglobulins on the direct bilirubin AU Beckman Coulter assay: the benefit of unsuspected diagnosis from spurious results
  16. Preliminary probe of quality indicators and quality specification in total testing process in 5753 laboratories in China
  17. Analytical and clinical evaluation of the new Fujirebio Lumipulse®G non-competitive assay for 25(OH)-vitamin D and three immunoassays for 25(OH)D in healthy subjects, osteoporotic patients, third trimester pregnant women, healthy African subjects, hemodialyzed and intensive care patients
  18. Patient-performed extraction of faecal calprotectin
  19. Comparison of the clinical utility of the Elia CTD Screen to indirect immunofluorescence on Hep-2 cells
  20. Reference Values and Biological Variations
  21. Sex-related differences in the association of ghrelin levels with obesity in adolescents
  22. Gestation specific reference intervals for thyroid function tests in pregnancy
  23. Cancer Diagnostics
  24. SOX17 promoter methylation in plasma circulating tumor DNA of patients with non-small cell lung cancer
  25. Dopamine concentration in blood platelets is elevated in patients with head and neck paragangliomas
  26. Letters to the Editor
  27. Cancer dynamics and the success of cancer screening programs
  28. Mother’s instinct – a rare case of multiple test interferences due to heterophile antibodies
  29. Sigma metric or defects per million opportunities (DPMO): the performance of clinical laboratories should be evaluated by the Sigma metrics at decimal level with DPMOs
  30. A national survey of preanalytical handling of oral glucose tolerance tests in pregnancy
  31. Updating pregnancy diabetes guidelines: is (y)our laboratory ready?
  32. Low serum bilirubin values are associated with pulmonary embolism in a case-control study
  33. Effect of Hb H on HbA1c measurements as measured by IFCC reference method and affinity HPLC
  34. Adipocytes in venipunctures cause falsely elevated S-100B serum values
  35. Earlier detection of sepsis by Candida parapsilosis using three-dimensional cytographic anomalies on the Mindray BC-6800 hematological analyzer
  36. Theranos phenomenon – part 4: Theranos at an International Conference
https://doi.org/10.1515/cclm-2015-0776
Keywords for this article
cell free DNA; circulating tumor DNA; DNA methylation; methylation specific PCR; non-small cell lung cancer; prognostic significance; SOX17

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. Laboratory analytical quality – the process continues
  4. Measurement uncertainty – a revised understanding of its calculation and use
  5. Review
  6. Substrate-zymography: a still worthwhile method for gelatinases analysis in biological samples
  7. Opinion Papers
  8. Is the combination of trueness and precision in one expression meaningful? On the use of total error and uncertainty in clinical chemistry
  9. The problem with total error models in establishing performance specifications and a simple remedy
  10. Measurement uncertainty for clinical laboratories – a revision of the concept
  11. Uncertainty in measurement and total error – are they so incompatible?
  12. General Clinical Chemistry and Laboratory Medicine
  13. Using the hazard ratio to evaluate allowable total error in predictive measurands
  14. Performance of electrolyte measurements assessed by a trueness verification program
  15. Analytical interference by monoclonal immunoglobulins on the direct bilirubin AU Beckman Coulter assay: the benefit of unsuspected diagnosis from spurious results
  16. Preliminary probe of quality indicators and quality specification in total testing process in 5753 laboratories in China
  17. Analytical and clinical evaluation of the new Fujirebio Lumipulse®G non-competitive assay for 25(OH)-vitamin D and three immunoassays for 25(OH)D in healthy subjects, osteoporotic patients, third trimester pregnant women, healthy African subjects, hemodialyzed and intensive care patients
  18. Patient-performed extraction of faecal calprotectin
  19. Comparison of the clinical utility of the Elia CTD Screen to indirect immunofluorescence on Hep-2 cells
  20. Reference Values and Biological Variations
  21. Sex-related differences in the association of ghrelin levels with obesity in adolescents
  22. Gestation specific reference intervals for thyroid function tests in pregnancy
  23. Cancer Diagnostics
  24. SOX17 promoter methylation in plasma circulating tumor DNA of patients with non-small cell lung cancer
  25. Dopamine concentration in blood platelets is elevated in patients with head and neck paragangliomas
  26. Letters to the Editor
  27. Cancer dynamics and the success of cancer screening programs
  28. Mother’s instinct – a rare case of multiple test interferences due to heterophile antibodies
  29. Sigma metric or defects per million opportunities (DPMO): the performance of clinical laboratories should be evaluated by the Sigma metrics at decimal level with DPMOs
  30. A national survey of preanalytical handling of oral glucose tolerance tests in pregnancy
  31. Updating pregnancy diabetes guidelines: is (y)our laboratory ready?
  32. Low serum bilirubin values are associated with pulmonary embolism in a case-control study
  33. Effect of Hb H on HbA1c measurements as measured by IFCC reference method and affinity HPLC
  34. Adipocytes in venipunctures cause falsely elevated S-100B serum values
  35. Earlier detection of sepsis by Candida parapsilosis using three-dimensional cytographic anomalies on the Mindray BC-6800 hematological analyzer
  36. Theranos phenomenon – part 4: Theranos at an International Conference
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 8.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/cclm-2015-0776/html
Scroll to top button