Cargando…

MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies

Plasma exosomal microRNAs (miRNAs) are considered as valid circulating biomarkers for cancer diagnosis and prognosis. Quantitative real-time polymerase chain reaction (qRT-PCR), the most commonly used technique to assess circulating miRNA levels, requires a normalization step involving uniformly exp...

Descripción completa

Detalles Bibliográficos
Autores principales: Damanti, Carlotta C., Gaffo, Enrico, Lovisa, Federica, Garbin, Anna, Di Battista, Piero, Gallingani, Ilaria, Tosato, Anna, Pillon, Marta, Carraro, Elisa, Mascarin, Maurizio, Elia, Caterina, Biffi, Alessandra, Bortoluzzi, Stefania, Mussolin, Lara
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7827902/
https://www.ncbi.nlm.nih.gov/pubmed/33429910
http://dx.doi.org/10.3390/cells10010101
_version_ 1783640880348921856
author Damanti, Carlotta C.
Gaffo, Enrico
Lovisa, Federica
Garbin, Anna
Di Battista, Piero
Gallingani, Ilaria
Tosato, Anna
Pillon, Marta
Carraro, Elisa
Mascarin, Maurizio
Elia, Caterina
Biffi, Alessandra
Bortoluzzi, Stefania
Mussolin, Lara
author_facet Damanti, Carlotta C.
Gaffo, Enrico
Lovisa, Federica
Garbin, Anna
Di Battista, Piero
Gallingani, Ilaria
Tosato, Anna
Pillon, Marta
Carraro, Elisa
Mascarin, Maurizio
Elia, Caterina
Biffi, Alessandra
Bortoluzzi, Stefania
Mussolin, Lara
author_sort Damanti, Carlotta C.
collection PubMed
description Plasma exosomal microRNAs (miRNAs) are considered as valid circulating biomarkers for cancer diagnosis and prognosis. Quantitative real-time polymerase chain reaction (qRT-PCR), the most commonly used technique to assess circulating miRNA levels, requires a normalization step involving uniformly expressed endogenous miRNAs. However, there is still no consensus on reference miRNAs for plasma exosomal miRNA abundance normalization. In this study, we identified a panel of miRNAs with stable abundance by analyzing public plasma exosome RNA-seq data and selected miR-486-5p, miR-26a-5p, miR-423-5p and miR191-5p as candidate normalizers. Next, we tested the abundance variation of these miRNAs by qRT-PCR in plasma exosomes of healthy donors and pediatric patients with anaplastic large cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma and mature B-cell acute lymphoblastic leukemia. MiR-486-5p and miR-26a-5p showed the most stable levels, both between healthy controls and patients and among the malignancies analyzed. In light of previous reports on miRNA stability in different exosome isolation methods, our data indicated that miR-26a-5p is a bona fide reference miRNA for qRT-PCR normalization to evaluate miRNA abundance from circulating plasma exosomes in studies of hematological malignancies.
format Online
Article
Text
id pubmed-7827902
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-78279022021-01-25 MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies Damanti, Carlotta C. Gaffo, Enrico Lovisa, Federica Garbin, Anna Di Battista, Piero Gallingani, Ilaria Tosato, Anna Pillon, Marta Carraro, Elisa Mascarin, Maurizio Elia, Caterina Biffi, Alessandra Bortoluzzi, Stefania Mussolin, Lara Cells Article Plasma exosomal microRNAs (miRNAs) are considered as valid circulating biomarkers for cancer diagnosis and prognosis. Quantitative real-time polymerase chain reaction (qRT-PCR), the most commonly used technique to assess circulating miRNA levels, requires a normalization step involving uniformly expressed endogenous miRNAs. However, there is still no consensus on reference miRNAs for plasma exosomal miRNA abundance normalization. In this study, we identified a panel of miRNAs with stable abundance by analyzing public plasma exosome RNA-seq data and selected miR-486-5p, miR-26a-5p, miR-423-5p and miR191-5p as candidate normalizers. Next, we tested the abundance variation of these miRNAs by qRT-PCR in plasma exosomes of healthy donors and pediatric patients with anaplastic large cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma and mature B-cell acute lymphoblastic leukemia. MiR-486-5p and miR-26a-5p showed the most stable levels, both between healthy controls and patients and among the malignancies analyzed. In light of previous reports on miRNA stability in different exosome isolation methods, our data indicated that miR-26a-5p is a bona fide reference miRNA for qRT-PCR normalization to evaluate miRNA abundance from circulating plasma exosomes in studies of hematological malignancies. MDPI 2021-01-08 /pmc/articles/PMC7827902/ /pubmed/33429910 http://dx.doi.org/10.3390/cells10010101 Text en © 2021 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
Damanti, Carlotta C.
Gaffo, Enrico
Lovisa, Federica
Garbin, Anna
Di Battista, Piero
Gallingani, Ilaria
Tosato, Anna
Pillon, Marta
Carraro, Elisa
Mascarin, Maurizio
Elia, Caterina
Biffi, Alessandra
Bortoluzzi, Stefania
Mussolin, Lara
MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies
title MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies
title_full MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies
title_fullStr MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies
title_full_unstemmed MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies
title_short MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies
title_sort mir-26a-5p as a reference to normalize microrna qrt-pcr levels in plasma exosomes of pediatric hematological malignancies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7827902/
https://www.ncbi.nlm.nih.gov/pubmed/33429910
http://dx.doi.org/10.3390/cells10010101
work_keys_str_mv AT damanticarlottac mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT gaffoenrico mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT lovisafederica mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT garbinanna mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT dibattistapiero mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT gallinganiilaria mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT tosatoanna mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT pillonmarta mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT carraroelisa mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT mascarinmaurizio mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT eliacaterina mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT biffialessandra mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT bortoluzzistefania mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies
AT mussolinlara mir26a5pasareferencetonormalizemicrornaqrtpcrlevelsinplasmaexosomesofpediatrichematologicalmalignancies