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Cancers 2016, 8(12), 112; doi:10.3390/cancers8120112

Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods

1
Exosomes Lab, CIC bioGUNE, CIBERehd, Derio 48160, Spain
2
Abcam, One Kendall Square, Cambridge, MA 02139, USA
3
Cancer Cell Signaling and Metabolism Lab, CIC bioGUNE, Derio 48160, Spain
4
Urology Service, Basurto University Hospital, Bilbao 48013, Spain
5
IKERBASQUE Basque Foundation for Science, Bilbao 48013, Spain
*
Authors to whom correspondence should be addressed.
Academic Editor: Samuel C. Mok
Received: 14 October 2016 / Revised: 25 November 2016 / Accepted: 6 December 2016 / Published: 10 December 2016
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Abstract

Urine extracellular vesicles are a valuable low-invasive source of information, especially for the cells of the genitourinary tract. In the search for biomarkers, different techniques have been developed to isolate and characterize the cargo of these vesicles. In the present work, we compare five of these different isolation methods (three commercial isolation kits, ultracentrifugation, and lectin-based purification) and perform miRNA profiling using a multiplex miRNA assay. The results showed high correlation through all isolation techniques, and 48 out of 68 miRNAs were detected above the detection limit at least 10 times. The results obtained by multiplex assay were validated through Taqman qPCR. In addition, using this technique combined with a clinically friendly extracellular vesicle (uEV)-enrichment method, we performed the analysis of selected miRNAs in urine from patients affected with bladder cancer, benign prostate hyperplasia, or prostate cancer. Importantly, we found that those miRNAs could be detected in almost 100% of the samples, and no significant differences were observed between groups. Our results support the feasibility of analyzing exosomes-associated miRNAs using a methodology that requires a small volume of urine and is compatible with a clinical environment and high-throughput analysis. View Full-Text
Keywords: extracellular vesicles; exosomes; urine; miRNA; isolation methods extracellular vesicles; exosomes; urine; miRNA; isolation methods
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MDPI and ACS Style

Royo, F.; Diwan, I.; Tackett, M.R.; Zuñiga, P.; Sanchez-Mosquera, P.; Loizaga-Iriarte, A.; Ugalde-Olano, A.; Lacasa, I.; Perez, A.; Unda, M.; Carracedo, A.; Falcon-Perez, J.M. Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods. Cancers 2016, 8, 112.

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