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

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...

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Autores principales: Royo, Felix, Diwan, Izzuddin, Tackett, Michael R., Zuñiga, Patricia, Sanchez-Mosquera, Pilar, Loizaga-Iriarte, Ana, Ugalde-Olano, Aitziber, Lacasa, Isabel, Perez, Amparo, Unda, Miguel, Carracedo, Arkaitz, Falcon-Perez, Juan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5187510/
https://www.ncbi.nlm.nih.gov/pubmed/27973407
http://dx.doi.org/10.3390/cancers8120112
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author Royo, Felix
Diwan, Izzuddin
Tackett, Michael R.
Zuñiga, Patricia
Sanchez-Mosquera, Pilar
Loizaga-Iriarte, Ana
Ugalde-Olano, Aitziber
Lacasa, Isabel
Perez, Amparo
Unda, Miguel
Carracedo, Arkaitz
Falcon-Perez, Juan M.
author_facet Royo, Felix
Diwan, Izzuddin
Tackett, Michael R.
Zuñiga, Patricia
Sanchez-Mosquera, Pilar
Loizaga-Iriarte, Ana
Ugalde-Olano, Aitziber
Lacasa, Isabel
Perez, Amparo
Unda, Miguel
Carracedo, Arkaitz
Falcon-Perez, Juan M.
author_sort Royo, Felix
collection PubMed
description 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.
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spelling pubmed-51875102016-12-30 Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods Royo, Felix Diwan, Izzuddin Tackett, Michael R. Zuñiga, Patricia Sanchez-Mosquera, Pilar Loizaga-Iriarte, Ana Ugalde-Olano, Aitziber Lacasa, Isabel Perez, Amparo Unda, Miguel Carracedo, Arkaitz Falcon-Perez, Juan M. Cancers (Basel) Article 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. MDPI 2016-12-10 /pmc/articles/PMC5187510/ /pubmed/27973407 http://dx.doi.org/10.3390/cancers8120112 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Royo, Felix
Diwan, Izzuddin
Tackett, Michael R.
Zuñiga, Patricia
Sanchez-Mosquera, Pilar
Loizaga-Iriarte, Ana
Ugalde-Olano, Aitziber
Lacasa, Isabel
Perez, Amparo
Unda, Miguel
Carracedo, Arkaitz
Falcon-Perez, Juan M.
Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods
title Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods
title_full Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods
title_fullStr Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods
title_full_unstemmed Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods
title_short Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods
title_sort comparative mirna analysis of urine extracellular vesicles isolated through five different methods
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5187510/
https://www.ncbi.nlm.nih.gov/pubmed/27973407
http://dx.doi.org/10.3390/cancers8120112
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