Proteome analysis of the urine has shown that urine contains disease-specific information for a variety of urogenital system disorders, including prostate cancer (PCa). The aim of this study was to determine the protein components of urine from PCa patients. Urine from 8 patients with clinically and histologically confirmed PCa was analyzed by conventional 2D PAGE. The MS identification of the most prominent 125 spots from the urine map revealed 45 distinct proteins. According to Gene Ontology, the identified proteins are involved in a variety of biological processes, majority of them are secreted (71%), and half of them are enzymes or transporters. Comparison with the normal urine proteome revealed 11 proteins distinctive for PCa. Using Ingenuity Pathways Analysis, we have found 3 proteins (E3 ubiquitin-protein ligase rififylin, tumor protein D52, and thymidine phosphorylase) associated with cellular growth and proliferation ( ). The top network of functional associations between 11 proteins was Cell Death and Survival, Cell-To-Cell Signaling and Interaction, and System Development and Function . In summary, we have created an initial proteomic map of PCa patient’s urine. The results from this study provide some leads to understand the molecular bases of prostate cancer. 1. Introduction Urine has become one of the most attractive biofluids in clinical proteomics because it can be obtained in large quantities, can be sampled noninvasively, and does not undergo significant proteolytic degradation compared with other biofluids [1]. The urine contains water, glucose, salt, and proteins derived from plasma or the urogenital tract. It can be viewed as modified ultrafiltrate of plasma combined with proteins derived from kidney and urinary tract, with protein concentration approximately 1000-fold lower than in plasma itself [2]. Even though the urinary proteome is much less complex than the plasma proteome, it contains high number of proteins. The urinary proteome has been studied by almost any proteomics technology. The first proteomic profiling of the normal urine was performed in 1979 using two-dimensional electrophoresis (2D) [3]. Afterwards, 2D, liquid chromatography (LC) and capillary electrophoresis (CE), all of them coupled to mass spectrometry (MS), have been used extensively in the proteomics definition of the urine. With the advent of the high throughput proteomics platforms consisting of 1D SDS-PAGE or LC coupled with high resolution mass spectrometers such as LTQ-FT and LTQ-Orbitrap, the number of detected proteins in healthy urine reached from
References
[1]
S. Decramer, A. G. de Peredo, B. Breuil et al., “Urine in clinical proteomics,” Molecular and Cellular Proteomics, vol. 7, no. 10, pp. 1850–1862, 2008.
[2]
G. L. Hortin and D. Sviridov, “Diagnostic potential for urinary proteomics,” Pharmacogenomics, vol. 8, no. 3, pp. 237–255, 2007.
[3]
N. G. Anderson, N. L. Anderson, and S. L. Tollaksen, “Proteins in human urine. I. Concentration and analysis by two-dimensional electrophoresis,” Clinical Chemistry, vol. 25, no. 7, pp. 1199–1210, 1979.
[4]
J. Adachi, C. Kumar, Y. Zhang, J. V. Olsen, and M. Mann, “The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins,” Genome Biology, vol. 7, no. 9, article R80, 2006.
[5]
Q. R. Li, K. X. Fan, R. X. Li et al., “A comprehensive and non-prefractionation on the protein level approach for the human urinary proteome: touching phosphorylation in urine,” Rapid Communications in Mass Spectrometry, vol. 24, no. 6, pp. 823–832, 2010.
[6]
A. Marimuthu, R. N. O'Meally, R. Chaerkady et al., “A comprehensive map of the human urinary proteome,” Journal of Proteome Research, vol. 10, no. 6, pp. 2734–2743, 2011.
[7]
G. Candiano, L. Santucci, A. Petretto et al., “2D-electrophoresis and the urine proteome map: where do we stand?” Journal of Proteomics, vol. 73, no. 5, pp. 829–844, 2010.
[8]
S. Magdeldin, S. Enany, Y. Yoshida, et al., “Basics and recent advances of two dimensional-polyacrylamide gel electrophoresis,” Clinical Proteomics, vol. 11, no. 1, article 16, 2014.
[9]
E. R. Suárez, J. Siwy, P. Zürbig, and H. Mischak, “Urine as a source for clinical proteome analysis: from discovery to clinical application,” Biochimica et Biophysica Acta, vol. 1844, no. 5, pp. 884–898, 2014.
[10]
A. Albalat, H. Mischak, and W. Mullen, “Clinical application of urinary proteomics/peptidomics,” Expert Review of Proteomics, vol. 8, no. 5, pp. 615–629, 2011.
[11]
M. M. Bradford, “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding,” Analytical Biochemistry, vol. 72, no. 1-2, pp. 248–254, 1976.
[12]
C. Gabay and I. Kushner, “Acute-phase proteins and other systemic responses to inflammation,” The New England Journal of Medicine, vol. 340, no. 6, pp. 448–454, 1999.
[13]
S. Akiyama, T. Furukawa, T. Sumizawa et al., “The role of thymidine phosphorylase, an angiogenic enzyme, in tumor progression,” Cancer Science, vol. 95, no. 11, pp. 851–857, 2004.
[14]
R. Honda, H. Tanaka, and H. Yasuda, “Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53,” The FEBS Letters, vol. 420, no. 1, pp. 25–27, 1997.
[15]
J. D. Oliner, J. A. Pietenpol, S. Thiagalingam, J. Gyuris, K. W. Kinzler, and B. Vogelstein, “Oncoprotein MDM2 conceals the activation domain of tumour suppressor p53,” Nature, vol. 362, no. 6423, pp. 857–860, 1993.
[16]
E. Meulmeester, Y. Pereg, Y. Shiloh, and A. G. Jochemsen, “ATM-mediated phosphorylations inhibit Mdmx/Mdm2 stabilization by HAUSP in favor of p53 activation,” Cell Cycle, vol. 4, no. 9, pp. 1166–1170, 2005.
[17]
P. Tennstedt, C. Bolch, G. Strobel, et al., “Patterns of TPD52 overexpression in multiple human solid tumor types analyzed by quantitative PCR,” International Journal of Oncology, vol. 44, no. 2, pp. 609–615, 2014.
[18]
M. J. Clague and S. Urbé, “Ubiquitin: same molecule, different degradation pathways,” Cell, vol. 143, no. 5, pp. 682–685, 2010.
[19]
D. Mukhopadhyay and H. Riezman, “Proteasome-independent functions of ubiquitin in endocytosis and signaling,” Science, vol. 315, no. 5809, pp. 201–205, 2007.
[20]
B. M. Kessler, “Ubiquitin–omics reveals novel networks and associations with human disease,” Current Opinion in Chemical Biology, vol. 17, no. 1, pp. 59–65, 2013.
[21]
Z. M. Bataineh and O. Habbal, “Immunoreactivity of ubiqitin in human prostate gland,” Neuroendocrinology Letters, vol. 27, no. 4, pp. 517–522, 2006.
[22]
C. A. Dinarello, “Proinflammatory cytokines,” Chest, vol. 118, no. 2, pp. 503–508, 2000.
[23]
L. Bertazza and S. Mocellin, “The dual role of tumor necrosis factor (TNF) in cancer biology,” Current Medicinal Chemistry, vol. 17, no. 29, pp. 3337–3352, 2010.
[24]
M. P. de Caestecker, E. Piek, and A. B. Roberts, “Role of transforming growth factor-β signaling in cancer,” Journal of the National Cancer Institute, vol. 92, no. 17, pp. 1388–1402, 2000.
[25]
M. R. Zaidi and G. Merlino, “The two faces of interferon-γ in cancer,” Clinical Cancer Research, vol. 17, no. 19, pp. 6118–6124, 2011.
