MicroRNAs in tumor samples and urinary extracellular vesicles as a putative diagnostic tool for muscle-invasive bladder cancer
The identification of biomarkers characterizing the invasive potential of bladder cancer could enhance the clinical management of individual patients and therefore improve prognosis. The aim of this study was to define a miRNA panel in tumor tissues as well as in urinary extracellular vesicles (EVs) for discriminating muscle-invasive bladder cancer (MIBC) from non-muscle-invasive bladder cancer (NMIBC).
miRNA expression was analyzed in 24 formalin-fixed, paraffin-embedded (FFPE) tumor samples by microarray analysis and was further validated by qRT-PCR in 56 FFPE tumor samples as well as in 37 urinary EV samples.
Microarray analysis revealed 63 miRNAs that were significantly differentially expressed (P < 0.05) between tissues from MIBC and NMIBC tumors. Five selected miRNAs (miR-146b-5p, miR-155-5p, miR-138-5p, miR-144-5p, and miR-200a-3p) were validated by qRT-PCR. The expression of all except miR-144-5p was significantly associated with high tumor grade. In urinary EVs, a different expression was verified for miR-146b-5p (P = 0.004) and miR-155-5p (P = 0.036), which exhibited significantly higher expression in urinary EVs from patients with MIBC.
miRNAs are promising biomarkers for the identification of invasive bladder carcinomas. Tissue samples as well as urinary EVs may serve as sources for miRNA analysis. This method, in addition to histopathology, could provide a new diagnostic tool and facilitate individual therapeutic decisions to select patients for early cystectomy.
KeywordsBladder cancer Muscle-invasive bladder cancer microRNA Extracellular vesicle Urine
We gratefully acknowledge Laura Witt, Alexander Vogt and Sebastian Hölters from our department for excellent technical support.
SB and KJ contributed to the study conception and design. MS, SB and KJ contributed to the patient assessment and recruitment and sample collection. AP and RB contributed to the pathological diagnosis. SB, JH, PM and PE contributed to the molecular biology analyses. SB, KJ, JH, PM, PE, JuH and MS contributed to the statistical analyses and data interpretation. SB, KJ, JuH and MS contributed to the manuscript drafting and intellectual input. SB, KJ, JuH and MS contributed to the writing of the final manuscript version. All authors read and approved the final manuscript.
Funding was received from Saarland University, Saarbrücken, Germany, for the microarray analysis of tumor tissues and partially for the RT-PCR validation of tumor tissues and EVs.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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 Helsinki Declaration and its later amendments or comparable ethical standards.
Human and animal rights statement
This article does not contain any studies with animals performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
- Babjuk M, Bohle A, Burger M, Capoun O, Cohen D, Comperat EM, Hernandez V, Kaasinen E, Palou J, Roupret M, van Rhijn BW, Shariat SF, Soukup V, Sylvester RJ, Zigeuner R (2017) EAU guidelines on non-muscle-invasive urothelial carcinoma of the bladder: update 2016. Eur Urol 71:447–461CrossRefGoogle Scholar
- 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. Can Res 69:4851–4860CrossRefGoogle Scholar
- Eble JN, Sauter G, Epstein JI, Sesterhenn IA (2004) WHO classification of tumours. Pathology and genetics of tumours of the urinary system and male genital organs. IARC Press, LyonGoogle Scholar
- Edge S, Byrd DR, Compton CC, Fritz AG, Greene F, Trotti A (2010) AJCC cancer staging handbook. Springer, New YorkGoogle Scholar
- Jiang X, Du L, Wang L, Li J, Liu Y, Zheng G, Qu A, Zhang X, Pan H, Yang Y, Wang C (2015) Serum microRNA expression signatures identified from genome-wide microRNA profiling serve as novel noninvasive biomarkers for diagnosis and recurrence of bladder cancer. Int J Cancer 136:854–862CrossRefGoogle Scholar
- Matsuzaki K, Fujita K, Jingushi K, Kawashima A, Ujike T, Nagahara A, Ueda Y, Tanigawa G, Yoshioka I, Ueda K, Hanayama R, Uemura M, Miyagawa Y, Tsujikawa K, Nonomura N (2017) MiR-21-5p in urinary extracellular vesicles is a novel biomarker of urothelial carcinoma. Oncotarget 8:24668–24678CrossRefGoogle Scholar
- 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 J Int Cancer 132:2479–2491CrossRefGoogle Scholar
- Robertson AG, Kim J, Al-Ahmadie H, Bellmunt J, Guo G, Cherniack AD, Hinoue T, Laird PW, Hoadley KA, Akbani R, Castro MAA, Gibb EA, Kanchi RS, Gordenin DA, Shukla SA, Sanchez-Vega F, Hansel DE, Czerniak BA, Reuter VE, Su X, de Sa Carvalho B, Chagas VS, Mungall KL, Sadeghi S, Pedamallu CS, Lu Y, Klimczak LJ, Zhang J, Choo C, Ojesina AI, Bullman S, Leraas KM, Lichtenberg TM, Wu CJ, Schultz N, Getz G, Meyerson M, Mills GB, McConkey DJ, Weinstein JN, Kwiatkowski DJ, Lerner SP (2017) Comprehensive molecular characterization of muscle-invasive bladder cancer. Cell 171(540–556):e525Google Scholar