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Assessment of nucleosides as putative tumor biomarkers in prostate cancer screening by CE–UV

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Abstract

Cancer is responsible for millions of deaths worldwide, but most base diseases may be cured if detected early. Screening tests may be used to identify early-stage malignant neoplasms. However, the major screening tool for prostate cancer, the prostate-specific antigen test, has unsuitable sensitivity. Since cancer cells may affect the pattern of consumption and excretion of nucleosides, such biomolecules are putative biomarkers that can be used for diagnosis and treatment evaluation. Using a previously validated method for the analysis of nucleosides in blood serum by capillary electrophoresis with UV–vis spectroscopy detection, we investigated 60 samples from healthy individuals and 42 samples from prostate cancer patients. The concentrations of nucleosides in both groups were compared and a multivariate partial least squares–discriminant analysis classification model was optimized for prediction of prostate cancer. The validation of the model with an independent sample set resulted in the correct classification of 82.4% of the samples, with sensitivity of 90.5% and specificity of 76.7%. A significant downregulation of 5-methyluridine and inosine was observed, which can be indicative of the carcinogenic process. Therefore, such analytes are potential candidates for prostate cancer screening.

Separation of the studied nucleosides and the internal standard 8-Bromoguanosine by CE-UV (a); classification of the external validation samples (30 from healthy volunteers and 21 from prostate cancer patients) by the developed Partial Least Square – Discriminant Analysis (PLS-DA) model with accuracy of 82.4% (b); Receiver Operating Characteristics (ROC) curve (c); and Variable Importance in the Projection (VIP) values for the studied nucleosides (d). A significant down-regulation of 5- methyluridine (5mU) and inosine (I) was observed, which can be indicative of the presence of prostate tumors.

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References

  1. World Health Organization (2014). Word Health Organization. http://www.who.int. Accessed 18 Mar 2014

  2. Ruddon RW. Cancer Biology. 4th ed. New York: Oxford University Press; 2007.

    Google Scholar 

  3. Greave M. Cancer: the evolutionary legacy. 1st ed. New York: Oxford University Press; 2000.

    Google Scholar 

  4. Naylor S. Biomarkers: current perspectives and future prospects. Expert Rev Mol Diagn. 2003;3:525–9.

    Article  Google Scholar 

  5. Atkinson AJ, Wayne AC, DeGruttola VG, et al. Biomarkers and surrogate endpoints: Preferred definitions and conceptual framework. Clin Pharmacol Ther. 2001;69:89–95.

    Article  Google Scholar 

  6. Ramautar R, Somsen GW, de Jong GJ. CE-MS for metabolomics: developments and applications in the period 2010-2012. Electrophoresis. 2013;34:86–98.

    Article  CAS  Google Scholar 

  7. Jain KK. The handbook of biomarkers. New York: Humana; 2010.

    Book  Google Scholar 

  8. Vaidya VS, Bonventre JV, editors. Biomarkers in medicine, drug discovery and enviromental health, 1st ed. Hoboken: Wiley; 2010.

    Google Scholar 

  9. Bloomfield VA. Nucleic acids: stuctures, properties and functions. Sausalito: University Science Books; 2000.

    Google Scholar 

  10. Schram KH. Urinary nucleosides. Mass Spectrom Rev. 1998;17:131–251.

    Article  CAS  Google Scholar 

  11. Markuszewski MJ, Struck W, Waszczuk-Jankowska M, Kaliszan R. Metabolomic approach for determination of urinary nucleosides as potential tumor markers using electromigration techniques. Electrophoresis. 2010;31:2300–10.

    Article  CAS  Google Scholar 

  12. Bullinger D, Fröhlich H, Klaus F, et al. Bioinformatical evaluation of modified nucleosides as biomedical markers in diagnosis of breast cancer. Anal Chim Acta. 2008;618:29–34.

    Article  CAS  Google Scholar 

  13. Cho SH, Jung BH, Lee SH, Lee WY, Kong G, Chung BC. Direct determination of nucleosides in the urine of patients with breast cancer using column-switching liquid chromatography-tandem mass spectrometry. Biomed Chromatogr. 2006;20:1229–36.

    Article  CAS  Google Scholar 

  14. Djukovic D, Baniasadi HR, Kc R, Hammoud Z, Raftery D. Targeted serum metabolite profiling of nucleosides in esophageal adenocarcinoma. Rapid Commun Mass Spectrom. 2010;24:3057–62.

    Article  CAS  Google Scholar 

  15. Feng B, Zheng MH, Zheng YF, Lu AG, Li JW, Wang ML, et al. Normal and modified urinary nucleosides represent novel biomarkers for colorectal cancer diagnosis and surgery monitoring. J Gastroenterol Hepatol. 2005;20:1913–9.

    Article  Google Scholar 

  16. Fischbein A, Sharma OK, Selikoff IJ, Borek E. Urinary excretion of modified nucleosides in patients with malignant mesothelioma. Cancer Res. 1983;43:2971–4.

    CAS  Google Scholar 

  17. García-Closas M, Malats N, Real FX, et al. Genetic variation in the nucleotide excision repair pathway and bladder cancer risk. Cancer Epidemiol Biomarkers Prev. 2006;15:536–42.

    Article  Google Scholar 

  18. Henneges C, Bullinger D, Fux R, et al. Prediction of breast cancer by profiling of urinary RNA metabolites using support vector machine-based feature selection. BMC Cancer. 2009;9:104–14.

    Article  Google Scholar 

  19. Hsu WY, Chen CJ, Huang YC, Tsai FJ, Bin JL, Lai CC. Urinary nucleosides as biomarkers of breast, colon, lung, and gastric cancer in Taiwanese. PLoS One. 2013;8, e81701.

    Article  Google Scholar 

  20. Jeng LB, Lo WY, Hsu WY, Lin WD, Lin CT, Lai CC, et al. Analysis of urinary nucleosides as helper tumor markers in hepatocellular carcinoma diagnosis. Rapid Commun Mass Spectrom. 2009;23:1543–9.

    Article  CAS  Google Scholar 

  21. Kim KR, La S, Kim A, Kim JH, Liebich HM. Capillary electrophoretic profiling and pattern recognition analysis of urinary nucleosides from uterine myoma and cervical cancer patients. J Chromatogr B. 2001;754:97–106.

    Article  CAS  Google Scholar 

  22. La S, Cho JH, Kim JH, Kim KR. Capillary electrophoretic profiling and pattern recognition analysis of urinary nucleosides from thyroid cancer patients. Anal Chim Acta. 2003;486:171–82.

    Article  CAS  Google Scholar 

  23. Mao Y, Zhao X, Wang S, Cheng Y. Urinary nucleosides based potential biomarker selection by support vector machine for bladder cancer recognition. Anal Chim Acta. 2007;598:34–40.

    Article  CAS  Google Scholar 

  24. Zheng YF, Kong HW, Xiong JH, Lv S, Xu GW. Clinical significance and prognostic value of urinary nucleosides in breast cancer patients. Clin Biochem. 2005;38:24–30.

    Article  CAS  Google Scholar 

  25. Zheng YF, Yang J, Zhao XJ, Feng B, Kong HW, Chen YJ, et al. Urinary nucleosides as biological markers for patients with colorectal cancer. World J Gastroenterol. 2005;11:3871–6.

    Article  CAS  Google Scholar 

  26. Thompson IM, Tangen CM, Kristal AR. Prostate-specific antigen: a misused and maligned prostate cancer biomarker. J Natl Cancer Inst. 2008;100:1487–8.

    Article  Google Scholar 

  27. Thompson IM, Ankerst DP, Chi C, Lucia S, Goodman PJ, Crowley JJ, et al. Operating characteristics of prostate-specific antigen in men with an initial PSA level of 3.0 ng/mL or lower. JAMA. 2005;294:66.

    Article  CAS  Google Scholar 

  28. Catalona WJ, Smith DS, Ornstein DK. Prostate cancer detection in men with serum PSA concentrations of 2.6 to 4.0 ng/mL and benign prostate examination. JAMA. 1997;277:1452.

    Article  CAS  Google Scholar 

  29. Brawer MK, Meyer GE, Letran JL, Bankson DD, Morris DL, Yeung KK, et al. Measurement of complexed PSA improves specificity for early detection of prostate cancer. Urology. 1998;52:372–8.

    Article  CAS  Google Scholar 

  30. Klein EA, Jones JS, editors. Management of prostate cancer. 3rd ed. Humana: New York; 2013.

    Google Scholar 

  31. Mydlo JH, Godec CJ, editors. Prostate cancer: science and clinical practice. 1st ed. Elsevier: London; 2003.

    Google Scholar 

  32. Woolf SH, Jonas S, Lawrence RS, editors. Health promotion and disease prevention in clinical practice. Philadelphia: Lippincott Williams & Wilkins; 1996.

    Google Scholar 

  33. Struck W, Waszczuk-Jankowska M, Kaliszan R, Markuszewski MJ. The state-of-the-art determination of urinary nucleosides using chromatographic techniques “hyphenated” with advanced bioinformatic methods. Anal Bioanal Chem. 2011;401:2039–50.

    Article  CAS  Google Scholar 

  34. Simionato AVC, Carrilho E, Maggi Tavares MF. CE-MS and related techniques as a valuable tool in tumor biomarkers research. Electrophoresis. 2010;31:1214–26.

    Article  CAS  Google Scholar 

  35. Buzatto AZ, de Sousa AC, Guedes SF, Cieslarová Z, Simionato AVC. Metabolomic investigation of human diseases biomarkers by CE and LC coupled to MS. Electrophoresis. 2014;35:1285–307.

    Article  CAS  Google Scholar 

  36. Buzatto AZ, Guedes SF, de Oliveira SM, Gallafrio JM, Simionato AVC. Higher detectability method for the analysis of nucleosides, putative tumor biomarkers, in blood serum samples by CE-UV with reversed EOF. Electrophoresis. 2015;36:2968–75.

    Article  CAS  Google Scholar 

  37. Brazilian National Health Surveillance Agency - ANVISA (2003) Guia de validação de métodos bioanalíticos, RE no 899, de 29 de maio de 2003.

  38. Food and Drug Administration (2001) Guidance for industry - bioanalytical method validation. US Department of Health and Human Services, Washington.

  39. European Medicines Agency. Guideline on validation of bioanalytical method validation. London: European Medicines Agency; 2009.

    Google Scholar 

  40. Jiang Y, Ma Y. A fast capillary electrophoresis method for separation and quantification of modified nucleosides in urinary samples. Anal Chem. 2009;81:6474–80.

    Article  CAS  Google Scholar 

  41. Szymańska E, Markuszewski MJ, Bodzioch K, Kaliszan R. Development and validation of urinary nucleosides and creatinine assay by capillary electrophoresis with solid phase extraction. J Pharm Biomed Anal. 2007;44:1118–26.

    Article  Google Scholar 

  42. Liebich HM, Xu GW, Di Stefano C, Lehmann R, Häring HU, Lu P, et al. Analysis of normal and modified nucleosides in urine by capillary electrophoresis. Chromatographia. 1997;45:396–401.

    Article  CAS  Google Scholar 

  43. Zheng YF, Xu GW, Liu DY, Xiong JH, Zhang PD, Zhang C, et al. Study of urinary nucleosides as biological marker in cancer patients analyzed by micellar electrokinetic capillary chromatography. Electrophoresis. 2002;23:4104–9.

    Article  CAS  Google Scholar 

  44. Paz N, Levanon EY, Amariglio N, et al (2007) Altered adenosine-to-inosine RNA editing in human cancer. Genome Res 1586–1595

  45. Grosjean H. Modification and editing of RNA: historical overview and important facts to remember. Top Curr Genet. 2005;12:1–22.

    Article  CAS  Google Scholar 

  46. Alseth I, Dalhus B, Bjørås M. Inosine in DNA and RNA. Curr Opin Genet Dev. 2014;26:116–23.

    Article  CAS  Google Scholar 

  47. Dominissini D, Moshitch-Moshkovitz S, Amariglio N, Rechavi G. Adenosine-to-inosine RNA editing meets cancer. Carcinogenesis. 2011;32:1569–77.

    Article  CAS  Google Scholar 

  48. Gallo A, Locatelli F. ADARs: allies or enemies? The importance of A-to-I RNA editing in human disease: from cancer to HIV-1. Biol Rev Camb Philos Soc. 2012;87:95–110.

    Article  Google Scholar 

  49. Akhavan-Niaki H, Samadani AA. DNA methylation and cancer development: molecular mechanism. Cell Biochem Biophys. 2013;67:501–13.

    Article  CAS  Google Scholar 

  50. Ehrlich M. DNA methylation in cancer: too much, but also too little. Oncogene. 2002;21:5400–13.

    Article  CAS  Google Scholar 

  51. Carmona FJ, Azuara D, Berenguer-Llergo A, et al. DNA methylation biomarkers for noninvasive diagnosis of colorectal cancer. Cancer Prev Res. 2013;6:656–65.

    Article  CAS  Google Scholar 

  52. Formosa A, Lena AM, Markert EK, et al. DNA methylation silences miR-132 in prostate cancer. Oncogene. 2013;32:127–34.

    Article  CAS  Google Scholar 

  53. Aran D, Hellman A. DNA Methylation of transcriptional enhancers and cancer predisposition. Cell. 2013;154:11–3.

    Article  CAS  Google Scholar 

  54. Aran D, Sabato S, Hellman A. DNA methylation of distal regulatory sites characterizes dysregulation of cancer genes. Genome Biol. 2013;14:R21.

    Article  Google Scholar 

  55. Worley B, Powers R. Multivariate analysis in metabolomics. Curr Metabolomics. 2012;1:92–107.

    Google Scholar 

  56. Ballabio D, Consonni V. Classification tools in chemistry. Part 1: linear models. PLS-DA. Anal. Methods. 2013;5:3790.

    CAS  Google Scholar 

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Acknowledgments

The authors thank the Clinics Hospital Blood Center (University of Campinas) and Laurione Candido de Oliveira for providing the serum samples from prostate cancer patients, Coordination for the Improvement of Higher Education Personnel (CAPES) for a scholarship to A.Z.B, the National Council for Scientific and Technological Development (CNPq) for financial support, and São Paulo Research Foundation (FAPESP) for financial support and a scholarship for M.O.S..

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Authors and Affiliations

  1. Department of Analytical Chemistry, Institute of Chemistry, University of Campinas, P.O. Box 6154, 13083-970, Campinas, São Paulo, Brazil

    Adriana Zardini Buzatto, Mariana de Oliveira Silva, Ronei Jesus Poppi & Ana Valéria Colnaghi Simionato

  2. National Institute of Science and Technology for Bioanalytics, Institute of Chemistry, University of Campinas, P.O. Box 6154, 13083-970, Campinas, São Paulo, Brazil

    Ronei Jesus Poppi & Ana Valéria Colnaghi Simionato

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  1. Adriana Zardini Buzatto
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Correspondence to Ana Valéria Colnaghi Simionato.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

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This study was funded by Coordination for the Improvement of Higher Education Personnel (CAPES), the National Council for Scientific and Technological Development (CNPq), and São Paulo Research Foundation (FAPESP).

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Buzatto, A.Z., de Oliveira Silva, M., Poppi, R.J. et al. Assessment of nucleosides as putative tumor biomarkers in prostate cancer screening by CE–UV. Anal Bioanal Chem 409, 3289–3297 (2017). https://doi.org/10.1007/s00216-017-0297-7

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