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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...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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 |
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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 |
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