Skip to main content
SpringerLink
Log in

Quantification of MicroRNAs in Urine-Derived Specimens

  • Protocol
  • First Online:
Urothelial Carcinoma

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1655))

Abstract

MicroRNAs are small noncoding RNAs which regulate the expression of genes involved in a multitude of cellular processes. Dysregulation of microRNAs and—in consequence—of the affected pathways is frequently observed in numerous pathologies including cancers. Therefore, tumor-related alterations in microRNA expression and function can reflect molecular processes of tumor onset and progression qualifying microRNAs as potential diagnostic and prognostic biomarkers.

In particular, microRNAs with differential expression in bladder cancer (BCa) might represent promising tools for noninvasive tumor detection in urine. This would be helpful not only for diagnostic and monitoring purposes but also for therapeutic decisions. Detection and quantification of BCa-associated microRNAs in urine can be performed using the cellular sediment, which also contains BCa cells, or in exosomes originating from those cells. Methods for isolation of exosomes from urine, extraction of total RNA from cells and exosomes as well as techniques for RNA quantification, reverse transcription, and qPCR-based quantification of microRNA expression levels are described herein.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Institutional subscriptions

References

  1. Carrington JC, Ambros V (2003) Role of microRNAs in plant and animal development. Science 301(5631):336–338. doi:10.1126/science.1085242

    Article  CAS  PubMed  Google Scholar 

  2. Mlcochova H, Hezova R, Stanik M, Slaby O (2014) Urine microRNAs as potential noninvasive biomarkers in urologic cancers. Urol Oncol 32(1):41 e41–41 e49. doi:10.1016/j.urolonc.2013.04.011

    Article  Google Scholar 

  3. Fendler A, Stephan C, Yousef GM, Jung K (2011) MicroRNAs as regulators of signal transduction in urological tumors. Clin Chem 57(7):954–968. doi:10.1373/clinchem.2010.157727

    Article  CAS  PubMed  Google Scholar 

  4. Schubert M, Junker K, Heinzelmann J (2016) Prognostic and predictive miRNA biomarkers in bladder, kidney and prostate cancer: where do we stand in biomarker development? J Cancer Res Clin Oncol 142(8):1673–1695. doi:10.1007/s00432-015-2089-9

    Article  CAS  PubMed  Google Scholar 

  5. Catto JW, Alcaraz A, Bjartell AS, De Vere WR, Evans CP, Fussel S, Hamdy FC, Kallioniemi O, Mengual L, Schlomm T, Visakorpi T (2011) MicroRNA in prostate, bladder, and kidney cancer: a systematic review. Eur Urol 59(5):671–681. doi:10.1016/j.eururo.2011.01.044

    Article  CAS  PubMed  Google Scholar 

  6. Ouyang H, Zhou Y, Zhang L, Shen G (2015) Diagnostic value of microRNAs for urologic cancers: a systematic review and meta-analysis. Medicine (Baltimore) 94(37):e1272. doi:10.1097/md.0000000000001272

    Article  CAS  Google Scholar 

  7. Canturk KM, Ozdemir M, Can C, Oner S, Emre R, Aslan H, Cilingir O, Ciftci E, Celayir FM, Aldemir O, Ozen M, Artan S (2014) Investigation of key miRNAs and target genes in bladder cancer using miRNA profiling and bioinformatic tools. Mol Biol Rep 41(12):8127–8135. doi:10.1007/s11033-014-3713-5

    Article  CAS  PubMed  Google Scholar 

  8. Dyrskjot L, Ostenfeld MS, Bramsen JB, Silahtaroglu AN, Lamy P, Ramanathan R, Fristrup N, Jensen JL, Andersen CL, Zieger K, Kauppinen S, Ulhoi BP, Kjems J, Borre M, Orntoft TF (2009) Genomic profiling of microRNAs in bladder cancer: miR-129 is associated with poor outcome and promotes cell death in vitro. Cancer Res 69(11):4851–4860. doi:10.1158/0008-5472.can-08-4043

    Article  CAS  PubMed  Google Scholar 

  9. Han Y, Chen J, Zhao X, Liang C, Wang Y, Sun L, Jiang Z, Zhang Z, Yang R, Li Z, Tang A, Li X, Ye J, Guan Z, Gui Y, Cai Z (2011) MicroRNA expression signatures of bladder cancer revealed by deep sequencing. PLoS One 6(3):e18286. doi:10.1371/journal.pone.0018286

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ichimi T, Enokida H, Okuno Y, Kunimoto R, Chiyomaru T, Kawamoto K, Kawahara K, Toki K, Kawakami K, Nishiyama K, Tsujimoto G, Nakagawa M, Seki N (2009) Identification of novel microRNA targets based on microRNA signatures in bladder cancer. Int J Cancer 125(2):345–352. doi:10.1002/ijc.24390

    Article  CAS  PubMed  Google Scholar 

  11. Itesako T, Seki N, Yoshino H, Chiyomaru T, Yamasaki T, Hidaka H, Yonezawa T, Nohata N, Kinoshita T, Nakagawa M, Enokida H (2014) The microRNA expression signature of bladder cancer by deep sequencing: the functional significance of the miR-195/497 cluster. PLoS One 9(2):e84311. doi:10.1371/journal.pone.0084311

    Article  PubMed  PubMed Central  Google Scholar 

  12. Zaravinos A, Radojicic J, Lambrou GI, Volanis D, Delakas D, Stathopoulos EN, Spandidos DA (2012) Expression of miRNAs involved in angiogenesis, tumor cell proliferation, tumor suppressor inhibition, epithelial-mesenchymal transition and activation of metastasis in bladder cancer. J Urol 188(2):615–623. doi:10.1016/j.juro.2012.03.122

    Article  CAS  PubMed  Google Scholar 

  13. Catto JW, Miah S, Owen HC, Bryant H, Myers K, Dudziec E, Larre S, Milo M, Rehman I, Rosario DJ, Di Martino E, Knowles MA, Meuth M, Harris AL, Hamdy FC (2009) Distinct microRNA alterations characterize high- and low-grade bladder cancer. Cancer Res 69(21):8472–8481. doi:10.1158/0008-5472.can-09-0744

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Dip N, Reis ST, Timoszczuk LS, Viana NI, Piantino CB, Morais DR, Moura CM, Abe DK, Silva IA, Srougi M, Dall'Oglio MF, Leite KR (2012) Stage, grade and behavior of bladder urothelial carcinoma defined by the microRNA expression profile. J Urol 188(5):1951–1956. doi:10.1016/j.juro.2012.07.004

    Article  PubMed  Google Scholar 

  15. Pignot G, Cizeron-Clairac G, Vacher S, Susini A, Tozlu S, Vieillefond A, Zerbib M, Lidereau R, Debre B, Amsellem-Ouazana D, Bieche I (2013) microRNA expression profile in a large series of bladder tumors: identification of a 3-miRNA signature associated with aggressiveness of muscle-invasive bladder cancer. Int J Cancer 132(11):2479–2491. doi:10.1002/ijc.27949

    Article  CAS  PubMed  Google Scholar 

  16. Sapre N, Macintyre G, Clarkson M, Naeem H, Cmero M, Kowalczyk A, Anderson PD, Costello AJ, Corcoran NM, Hovens CM (2016) A urinary microRNA signature can predict the presence of bladder urothelial carcinoma in patients undergoing surveillance. Br J Cancer 114(4):454–462. doi:10.1038/bjc.2015.472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Veerla S, Lindgren D, Kvist A, Frigyesi A, Staaf J, Persson H, Liedberg F, Chebil G, Gudjonsson S, Borg A, Mansson W, Rovira C, Hoglund M (2009) MiRNA expression in urothelial carcinomas: important roles of miR-10a, miR-222, miR-125b, miR-7 and miR-452 for tumor stage and metastasis, and frequent homozygous losses of miR-31. Int J Cancer 124(9):2236–2242. doi:10.1002/ijc.24183

    Article  CAS  PubMed  Google Scholar 

  18. Zhou H, Tang K, Xiao H, Zeng J, Guan W, Guo X, Xu H, Ye Z (2015) A panel of eight-miRNA signature as a potential biomarker for predicting survival in bladder cancer. J Exp Clin Cancer Res 34:53. doi:10.1186/s13046-015-0167-0

    Article  PubMed  PubMed Central  Google Scholar 

  19. Huang X, Liang M, Dittmar R, Wang L (2013) Extracellular microRNAs in urologic malignancies: chances and challenges. Int J Mol Sci 14(7):14785–14799. doi:10.3390/ijms140714785

    Article  PubMed  PubMed Central  Google Scholar 

  20. Armstrong DA, Green BB, Seigne JD, Schned AR, Marsit CJ (2015) MicroRNA molecular profiling from matched tumor and bio-fluids in bladder cancer. Mol Cancer 14:194. doi:10.1186/s12943-015-0466-2

    Article  PubMed  PubMed Central  Google Scholar 

  21. Eissa S, Habib H, Ali E, Kotb Y (2015) Evaluation of urinary miRNA-96 as a potential biomarker for bladder cancer diagnosis. Med Oncol 32(1):413. doi:10.1007/s12032-014-0413-x

    Article  PubMed  Google Scholar 

  22. Eissa S, Matboli M, Essawy NO, Kotb YM (2015) Integrative functional genetic-epigenetic approach for selecting genes as urine biomarkers for bladder cancer diagnosis. Tumour Biol 36(12):9545–9552. doi:10.1007/s13277-015-3722-6

    Article  CAS  PubMed  Google Scholar 

  23. Eissa S, Matboli M, Hegazy MG, Kotb YM, Essawy NO (2015) Evaluation of urinary microRNA panel in bladder cancer diagnosis: relation to bilharziasis. Transl Res 165(6):731–739. doi:10.1016/j.trsl.2014.12.008

    Article  CAS  PubMed  Google Scholar 

  24. Hanke M, Hoefig K, Merz H, Feller AC, Kausch I, Jocham D, Warnecke JM, Sczakiel G (2010) A robust methodology to study urine microRNA as tumor marker: microRNA-126 and microRNA-182 are related to urinary bladder cancer. Urol Oncol 28(6):655–661. doi:10.1016/j.urolonc.2009.01.027

    Article  CAS  PubMed  Google Scholar 

  25. Mengual L, Lozano JJ, Ingelmo-Torres M, Gazquez C, Ribal MJ, Alcaraz A (2013) Using microRNA profiling in urine samples to develop a non-invasive test for bladder cancer. Int J Cancer 133(11):2631–2641. doi:10.1002/ijc.28274

    CAS  PubMed  Google Scholar 

  26. Sasaki H, Yoshiike M, Nozawa S, Usuba W, Katsuoka Y, Aida K, Kitajima K, Kudo H, Hoshikawa M, Yoshioka Y, Kosaka N, Ochiya T, Chikaraishi T (2016) Expression level of urinary microRNA-146a-5p is increased in patients with bladder bancer and decreased in those after transurethral resection. Clin Genitourin Cancer 14(5):e493–e499. doi:10.1016/j.clgc.2016.04.002

    Article  PubMed  Google Scholar 

  27. Snowdon J, Boag S, Feilotter H, Izard J, Siemens DR (2013) A pilot study of urinary microRNA as a biomarker for urothelial cancer. Can Urol Assoc J 7(1–2):28–32. doi:10.5489/cuaj.11115

    Article  PubMed  PubMed Central  Google Scholar 

  28. Yamada Y, Enokida H, Kojima S, Kawakami K, Chiyomaru T, Tatarano S, Yoshino H, Kawahara K, Nishiyama K, Seki N, Nakagawa M (2011) MiR-96 and miR-183 detection in urine serve as potential tumor markers of urothelial carcinoma: correlation with stage and grade, and comparison with urinary cytology. Cancer Sci 102(3):522–529. doi:10.1111/j.1349-7006.2010.01816.x

    Article  CAS  PubMed  Google Scholar 

  29. Zhang DZ, Lau KM, Chan ES, Wang G, Szeto CC, Wong K, Choy RK, Ng CF (2014) Cell-free urinary microRNA-99a and microRNA-125b are diagnostic markers for the non-invasive screening of bladder cancer. PLoS One 9(7):e100793. doi:10.1371/journal.pone.0100793

    Article  PubMed  PubMed Central  Google Scholar 

  30. Mengual L, Ribal MJ, Lozano JJ, Ingelmo-Torres M, Burset M, Fernandez PL, Alcaraz A (2014) Validation study of a noninvasive urine test for diagnosis and prognosis assessment of bladder cancer: evidence for improved models. J Urol 191(1):261–269. doi:10.1016/j.juro.2013.06.083

    Article  PubMed  Google Scholar 

  31. Puerta-Gil P, Garcia-Baquero R, Jia AY, Ocana S, Alvarez-Mugica M, Alvarez-Ossorio JL, Cordon-Cardo C, Cava F, Sanchez-Carbayo M (2012) miR-143, miR-222, and miR-452 are useful as tumor stratification and noninvasive diagnostic biomarkers for bladder cancer. Am J Pathol 180(5):1808–1815. doi:10.1016/j.ajpath.2012.01.034

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Urology, Technische Universität Dresden, 01307, Dresden, Germany

    Susanne Fuessel, Andrea Lohse-Fischer, Dana Vu Van, Karsten Salomo, Kati Erdmann & Manfred P. Wirth

Authors
  1. Susanne Fuessel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrea Lohse-Fischer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dana Vu Van
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Karsten Salomo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kati Erdmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Manfred P. Wirth
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Susanne Fuessel .

Editor information

Editors and Affiliations

  1. Department of Urology, Heinrich Heine University, Düsseldorf, Germany

    Wolfgang A Schulz

  2. Department of Urology, Heinrich Heine University, Düsseldorf, Germany

    Michèle J. Hoffmann

  3. Department of Urology, Heinrich Heine University, Düsseldorf, Germany

    Günter Niegisch

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Fuessel, S., Lohse-Fischer, A., Vu Van, D., Salomo, K., Erdmann, K., Wirth, M.P. (2018). Quantification of MicroRNAs in Urine-Derived Specimens. In: Schulz, W., Hoffmann, M., Niegisch, G. (eds) Urothelial Carcinoma. Methods in Molecular Biology, vol 1655. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7234-0_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7234-0_16

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7233-3

  • Online ISBN: 978-1-4939-7234-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation