Abstract
Various microRNAs from the miR-371-3 and miR-302a–d clusters have recently been proposed as markers for testicular germ cell tumours. Upregulation of these miRNAs has been found in both the tissue and serum of testicular cancer patients, but they have never been studied in human seminal plasma. The aim of this study was, therefore, to assess the differences in the expression of miR-371-3 and miR-302a–d between the seminal plasma and serum of testicular cancer patients, and to identify new potential testicular cancer markers in seminal plasma. We investigated the serum and seminal plasma of 28 pre-orchiectomy patients subsequently diagnosed with testicular cancer, the seminal plasma of another 20 patients 30 days post-orchiectomy and a control group consisting of 28 cancer-free subjects attending our centre for an andrological check-up. Serum microRNA expression was analysed using RT-qPCR. TaqMan Array Card 3.0 platform was used for microRNA profiling in the seminal plasma of cancer patients. Results for both miR-371-3 and the miR-302 cluster in the serum of testicular cancer patients were in line with literature reports, while miR-371and miR-372 expression in seminal plasma showed the opposite trend to serum. On array analysis, 37 miRNAs were differentially expressed in the seminal plasma of cancer patients, and the upregulated miR-142 and the downregulated miR-34b were validated using RT-qPCR. Our study investigated the expression of miRNAs in the seminal plasma of patients with testicular cancer for the first time. Unlike in serum, miR-371-3 cannot be considered as markers in seminal plasma, whereas miR-142 levels in seminal plasma may be a potential marker for testicular cancer.
Similar content being viewed by others
References
J.W. Oosterhuis, L.H. Looijenga, Testicular germ-cell tumours in a broader perspective. Nat. Rev. Cancer. 5, 210–22 (2005)
C. Bosetti, P. Bertuccio, L. Chatenoud, E. Negri, C. La Vecchia, F. Levi, Trends in mortality from urologic cancers in Europe, 1970–2008. Eur. Urol. 60, 1–15 (2011)
F. Bray, L. Richiardi, A. Ekbom, D. Forman, E. Pukkala, M. Cuninkova, H. Møller, Do testicular seminoma and nonseminoma share the same etiology? Evidence from an age-period-cohort analysis of incidence trends in eight European countries. Cancer. Epidemiol. Biomark. Prev. 15, 652–658 (2006)
J. Beyer, P. Albers, R. Altena, J. Aparicio, C. Bokemeyer, J. Busch, R. Cathomas, E. Cavallin-Stahl, N.W. Clarke, J. Claßen, G. Cohn-Cedermark, A.A. Dahl, G. Daugaard, U. De Giorgi, M. De Santis, M. De Wit, R. De Wit, K.P. Dieckmann, M. Fenner, K. Fizazi, A. Flechon, S.D. Fossa, J.R. Germá Lluch, J.A. Gietema, S. Gillessen, A. Giwercman, J.T. Hartmann, A. Heidenreich, M. Hentrich, F. Honecker, A. Horwich, R.A. Huddart, S. Kliesch, C. Kollmannsberger, S. Krege, M.P. Laguna, L.H.J. Looijenga, J.P. Lorch ALotz, F. Mayer, A. Necchi, N. Nicolai, J. Nuver, K. Oechsle, J. Oldenburg, J.W. Oosterhuis, T. Powles, E. Rajpert-De Meyts, O. Rick, G. Rosti, R. Salvioni, M. Schrader, S. Schweyer, F. Sedlmayer, A. Sohaib, R. Souchon, T. Tandstad, C. Winter, C. Wittekind, Maintaining success, reducing treatment burden, focusing on survivorship: highlights from the third European consensus conference on diagnosis and treatment of germ-cell cancer. Ann. Oncol. 24, 878–888 (2013)
H.S. Haugnes, G.J. Bosl, H. Boer, J.A. Gietema, M. Brydøy, J. Oldenburg, A.A. Dahl, R.M. Bremnes, S.D. Fosså, Long-term and late effects of germ cell testicular cancer treatment and implications for follow-up. J. Clin. Oncol. 20, 3752–3763 (2012)
A. Horwich, J. Shipley, R. Huddart, Testicular germ cell cancer. Lancet. 29, 1398 (2006)
L.J. Barlow, G.M. Badalato, J.M. McKiernan, Serum tumor markers in the evaluation of male germ cell tumors. Nat. Rev. Urol. 7, 610–617 (2010)
V.N. Kim, J. Han, M.C. Siomi, Biogenesis of small RNAs in animals. Nat. Rev. Mol. Cell. Biol. 10, 126–139 (2009)
E. Rajpert-De Meyts, J.E. Nielsen, N.E. Skakkebaek, K. Almstrup, Diagnostic markers for germ cell neoplasms: from placental-like alkaline phosphatase to micro-RNAs. Folia Histochem. Cytobiol. 53, 177–188 (2015)
M.A. Rijlaarsdam, T. van Agthoven, A.J. Gillis, S. Patel, K. Hayashibara, K.Y. Lee, L.H. Looijenga, Identification of known and novel germ cell cancer-specific (embryonic) miRs in serum by high-throughput profiling. Andrology 3, 85–91 (2015)
K.P. Dieckmann, M. Spiekermann, T. Balks, I. Flor, T. Löning, J. Bullerdiek, G. Belge, MicroRNAs miR-371-3 in serum as diagnostic tools in the management of testicular germ cell tumours. Br. J. Cancer. 6, 1754–1760 (2012)
A.J. Gillis, H.J. Stoop, R. Hersmus, J.W. Oosterhuis, Y. Sun, C. Chen, S. Guenther, J. Sherlock, I. Veltman, J. Baeten, P.J. van der Spek, P. de Alarcon, L.H. Looijenga, High-throughput microRNAome analysis in human germ cell tumours. J. Pathol. 213, 319–328 (2007)
G. Belge, K.P. Dieckmann, M. Spiekermann, T. Balks, J. Bullerdiek, Serum levels of microRNAs miR-371-3: a novel class of serum biomarkers for testicular germ cell tumors? Eur. Urol. 61, 1068–1069 (2012)
I. Syring, J. Bartels, S. Holdenrieder, G. Kristiansen, S.C. Müller, J. Ellinger, Circulating serum miRNA (miR-367-3p, miR-371a-3p, miR-372-3p and miR-373-3p) as biomarkers in patients with testicular germ cell cancer. J. Urol. 193, 331–337 (2015)
C. Wang, C. Yang, X. Chen, B. Yao, C. Yang, C. Zhu, L. Li, J. Wang, X. Li, Y. Shao, Y. Liu, J. Ji, J. Zhang, K. Zen, C.Y. Zhang, C. Zhang, Altered profile of seminal plasma microRNAs in the molecular diagnosis of male infertility. Clin. Chem. 57, 1722–1731 (2011)
W. Wu, Z. Hu, Y. Qin, J. Dong, J. Dai, C. Lu, W. Zhang, H. Shen, Y. Xia, X. Wang, Seminal plasma microRNAs: potential biomarkers for spermatogenesis status. Mol. Hum. Reprod. 18, 489–497 (2012)
S.C. McIver, S.D. Roman, B. Nixon, E.A. McLaughlin, miRNA and mammalian male germ cells. Hum. Reprod. Update 18, 44–59 (2012)
M. Abu-Halima, C. Backes, P. Leidinger, A. Keller, A.M. Lubbad, M. Hammadeh, E. Meese, MicroRNA expression profiles in human testicular tissues of infertile men with different histopathologic patterns. Fertil. Steril. 101, 78–86 (2014)
L.A. Selth, M.J. Roberts, C.W. Chow, V.R. Marshall, S.A. Doi, A.D. Vincent, L.M. Butler, M.F. Lavin, W.D. Tilley, R.A. Gardiner, Human seminal fluid as a source of prostate cancer-specific microRNA biomarkers. Endocr. Relat. Cancer. 21, L17–21 (2014)
L.O. Reis, T.C. Pereira, I. Lopes-Cendes, U. Ferreira, MicroRNAs: a new paradigm on molecular urological oncology. Urology. 76, 521–527 (2010)
J.A. Weber, D.H. Baxter, S. Zhang, D.Y. Huang, K.H. Huang, M.J. Lee, D.J. Galas, K. Wang, The microRNA spectrum in 12 body fluids. Clin. Chem. 56, 1733–1741 (2010)
K.J. Livak, T.D. Schmittgen, Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25, 402–8 (2001)
J. Brennecke, D.R. Hipfner, A. Stark, R.B. Russell, S.M. Cohen, Bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113, 25–36 (2003)
Y.S. Lee, H.K. Kim, S. Chung, K.S. Kim, A. Dutta, Depletion of human micro-RNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the down-regulation of putative targets during differentiation. J. Biol. Chem. 280, 16635–16641 (2005)
R. Yi, M.N. Poy, M. Stoffel, E. Fuchs, A skin microRNA promotes differentiation by repressing ’stemness’. Nature 452, 225–229 (2008)
C.Z. Chen, L. Li, H.F. Lodish, D.P. Bartel, MicroRNAs modulate hematopoietic lineage differentiation. Science 2, 83–86 (2004)
V. Ambros, MicroRNA pathways in flies and worms: growth, death, fat, stress, and timing. Cell 113, 673–676 (2003)
P. Xu, S.Y. Vernooy, M. Guo, B.A. Hay, The Drosophila microRNA Mir-14 suppresses cell death and is required for normal fat metabolism. Curr. Biol. 29, 790–795 (2003)
A. Esquela-Kerscher, F.J. Slack, Oncomirs. microRNAs with a role in cancer. Nat. Rev. Cancer. 6, 259–269 (2006)
N. Kotaja, MicroRNAs and spermatogenesis. Fertil. Steril. 101, 1552–1562 (2014)
M.V. Iorio, C.M. Croce, microRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. EMBO Mol. Med. 4, 143–59 (2012)
L. Wang, C. Xu, Role of microRNAs in mammalian spermatogenesis and testicular germ cell tumors. Reproduction 149, R127–137 (2015)
Y. He, J. Lin, D. Kong, M. Huang, C. Xu, T.K. Kim, A. Etheridge, Y. Luo, Y. Ding, K. Wang, Current state of circulating MicroRNAs as cancer biomarkers. Clin. Chem. 61, 1138–1155 (2015)
S. Dimmeler, A.M. Zeiher, Circulating microRNAs: novel biomarkers for cardiovascular diseases? Eur. Heart J. 31, 2705–2707 (2010)
A. Zampetaki, S. Kiechl, I. Drozdov, P. Willeit, U. Mayr, M. Prokopi, A. Mayr, S. Weger, F. Oberhollenzer, E. Bonora, A. Shah, J. Willeit, M. Mayr, Plasma microRNA profiling reveals loss of endothelial miR-126 and other microRNAs in type 2 diabetes. Circ. Res. 17, 810–817 (2010)
K. Wang, S. Zhang, B. Marzolf, P. Troisch, A. Brightman, Z. Hu, L.E. Hood, D.J. Galas, Circulating microRNAs, potential biomarkers for drug-induced liver injury. Proc. Natl. Acad. Sci. USA 17, 4402–4407 (2009)
J. Li, X. Liang, Z. Chen, Improving the embryo implantation via novel molecular targets. Curr. Drug. Targets. 14, 864–871 (2013)
P.M. Voorhoeve, C. le Sage, M. Schrier, A.J. Gillis, H. Stoop, R. Nagel, Y.P. Liu, J. van Duijse, J. Drost, A. Griekspoor, E. Zlotorynski, N. Yabuta, G. De Vita, H. Nojima, L.H. Looijenga, R. Agami, A genetic screen implicates miRNA-372 and miRNA-373 as oncogenes in testicular germ cell tumors. Cell 24, 1169–1181 (2006)
R.D. Palmer, M.J. Murray, H.K. Saini, S. van Dongen, C. Abreu-Goodger, B. Muralidhar, M.R. Pett, C.M. Thornton, J.C. Nicholson, A.J. Enright, N. Coleman, Children’s cancer and leukaemia group malignant germ cell tumors display common microRNA profiles resulting in global changes in expression of messenger RNA targets. Cancer Res. 1, 2911–2923 (2010)
M.J. Murray, D.J. Halsall, C.E. Hook, D.M. Williams, J.C. Nicholson, N. Coleman, Identification of microRNAs from the miR-371~373 and miR-302 clusters as potential serum biomarkers of malignant germ cell tumors. Am. J. Clin. Pathol. 135, 119–125 (2011)
A.J. Gillis, M.A. Rijlaarsdam, R. Eini, L.C. Dorssers, K. Biermann, M.J. Murray, J.C. Nicholson, N. Coleman, K.P. Dieckmann, G. Belge, J. Bullerdiek, T. Xu, N. Bernard, L.H. Looijenga, Targeted serum miRNA (TSmiR) test for diagnosis and follow-up of (testicular) germ cell cancer patients: a proof of principle. Mol. Oncol. 7, 1083–1092 (2013)
M. Spiekermann, G. Belge, N. Winter, R. Ikogho, T. Balks, J. Bullerdiek, K.P. Dieckmann, MicroRNA miR-371a-3p in serum of patients with germ cell tumours: evaluations for establishing a serum biomarker. Andrology 3, 78–84 (2015)
Y. Chu, Y. Ouyang, F. Wang, A. Zheng, L. Bai, L. Han, Y. Chen, H. Wang, MicroRNA-590 promotes cervical cancer cell growth and invasion by targeting CHL1. J. Cell. Biochem. 115, 847–853 (2014)
F. Lian, Y. Cui, C. Zhou, K. Gao, L. Wu, Identification of a plasma four-microRNA panel as potential noninvasive biomarker for osteosarcoma. PLoS One. 16, e0121499 (2015)
J.L. Park, M. Kim, K.S. Song, S.Y. Kim, Y.S. Kim, Cell-Free miR-27a, a potential diagnostic and prognostic biomarker for gastric cancer. Genom. Inform 13, 70–75 (2015)
M. Bellon, Y. Lepelletier, O. Hermine, C. Nicot, Deregulation of microRNA involved in hematopoiesis and the immune response in HTLV-I adult T-cell leukemia. Blood. 14, 4914–4917 (2009)
A.B. Hu, M. Lenarduzzi, T. Krushel, L. Waldron, M. Pintilie, W. Shi, B. Perez-Ordonez, I. Jurisica, B. O’Sullivan, J. Waldron, P. Gullane, B. Cummings, F.F. Liu, Comprehensive MicroRNA profiling for head and neck squamous cell carcinomas. Clin. Cancer Res. 15, 1129–1139 (2010)
R.J. Lin, D.W. Xiao, L.D. Liao, T. Chen, Z.F. Xie, W.Z. Huang, W.S. Wang, T.F. Jiang, B.L. Wu, E.M. Li, L.Y. Xu, MiR-142-3p as a potential prognostic biomarker for esophageal squamous cell carcinoma. J. Surg. Oncol. 105, 175–182 (2012)
S. Kaduthanam, S. Gade, M. Meister, J.C. Brase, M. Johannes, H. Dienemann, A. Warth, P.A. Schnabel, F.J. Herth, H. Sültmann, T. Muley, R. Kuner, Serum miR-142-3p is associated with early relapse in operable lung adenocarcinoma patients. Lung Cancer. 80, 223–227 (2013)
L. Wu, C. Cai, X. Wang, M. Liu, X. Li, H. Tang, MicroRNA-142-3p, a new regulator of RAC1, suppresses the migration and invasion of hepatocellular carcinoma cells. FEBS Lett. 6, 1322–1330 (2011)
T. Isobe, S. Hisamori, D.J. Hogan, M. Zabala, D.G. Hendrickson, P. Dalerba, S. Cai, F. Scheeren, A.H. Kuo, S.S. Sikandar, J.S. Lam, D. Qian, F.M. Dirbas, G. Somlo, K. Lao, P.O. Brown, M.F. Clarke, Y. Shimono, miR-142 regulates the tumorigenicity of human breast cancer stem cells through the canonical WNT signaling pathway. eLife 3, e01977 (2014)
H. Liu, Y. Yang, L. Zhang, R. Liang, R.S. Ge, Y. Zhang, Q. Zhang, Q. Xiang, Y. Huang, Z. Su, Basic fibroblast growth factor promotes stem Leydig cell development and inhibits LH-stimulated androgen production by regulating microRNA expression. J. Steroid Biochem. Mol. Biol. 144, 483–491 (2014)
B. Huang, J. Zhao, Z. Lei, S. Shen, D. Li, G.X. Shen, G.M. Zhang, Z.H. Feng, miR-142-3p restricts cAMP production in CD4+ CD25- T cells and CD4+ CD25+ TREG cells by targeting AC9 mRNA. EMBO Rep. 10, 180–185 (2009)
D.M. Stocco, X. Wang, Y. Jo, P.R. Manna, Multiple signaling pathways regulating steroidogenesis and steroidogenic acute regulatory protein expression: more complicated than we thought. Mol. Endocrinol. 19, 2647–2659 (2005)
K. Tanaka, K. Kondo, K. Kitajima, M. Muraoka, A. Nozawa, T. Hara, Tumor-suppressive function of protein-tyrosine phosphatase non-receptor type 23 in testicular germ cell tumors is lost upon overexpression of miR142-3p microRNA. J. Biol. Chem. 16, 23990–23999 (2013)
H. Hermeking, The miR-34 family in cancer and apoptosis. Cell Death Differ. 17, 193–199 (2010)
S. Majid, A.A. Dar, S. Saini, V. Shahryari, S. Arora, M.S. Zaman, I. Chang, S. Yamamura, Y. Tanaka, T. Chiyomaru, G. Deng, R. Dahiya, miRNA-34b inhibits prostate cancer through demethylation, active chromatin modifications, and AKT pathways. Clin. Cancer Res. 1, 73–84 (2013)
M. Rokavek, H. Li, L. Jiang, H. Hermeking, The p53/miR-34 axis in development and disease. J. Mol. Cell Biol. 6, 214–230 (2014)
L. Smorag, Y. Zheng, J. Nolte, U. Zechner, W. Engel, D.V. Pantakani, MicroRNA signature in various cell types of mouse spermatogenesis: evidence for stage-specifically expressed miRNA-221, -203 and -34b-5p mediated spermatogenesis regulation. Biol. Cell. 104, 677–692 (2012)
Acknowledgments
The authors wish to thank Marie-Hélène Hayles for her assistance in the English translation of the manuscript.
Funding
This work was supported by a grant from the Italian Ministry of Education and Research (MIUR-PRIN) and the University of Rome “La Sapienza” Faculty of Medicine.
Author contributions
L.G., D.P.: Conception and design of the work. M.P., G.C.: Carried out the molecular genetic studies. L.G., D.P.: Drafting the article. L.G.: Final approval of the version to be published. M.P., F.P., G.C.: Acquisition and analysis of data. F.L., A.L., F.P.: Revising the paper critically. All authors read and approved the final manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Ethical approval
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.
Rights and permissions
About this article
Cite this article
Pelloni, M., Coltrinari, G., Paoli, D. et al. Differential expression of miRNAs in the seminal plasma and serum of testicular cancer patients. Endocrine 57, 518–527 (2017). https://doi.org/10.1007/s12020-016-1150-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12020-016-1150-z