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Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients
Background: Circulating microRNA (miRNA) analysis is a growing research field. However, it usually requires an endogenous control or housekeeping (HK) in order to normalize expression of specific miRNAs throughout different samples. Unfortunately, no adequate HK for circulating miRNA analysis is sti...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022916/ https://www.ncbi.nlm.nih.gov/pubmed/31877644 http://dx.doi.org/10.3390/biom10010016 |
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author | Duran-Sanchon, Saray Vila-Navarro, Elena Marcuello, Maria Lozano, Juan José Muñoz, Jenifer Cubiella, Joaquín Diez, Maria Soledad Bujanda, Luis Lanas, Angel Jover, Rodrigo Hernández, Vicent Quintero, Enrique Herreros-Villanueva, Marta Martín, Ana Carmen Pérez-Palacios, Rosa Arroyo, Rocio Castells, Antoni Gironella, Meritxell |
author_facet | Duran-Sanchon, Saray Vila-Navarro, Elena Marcuello, Maria Lozano, Juan José Muñoz, Jenifer Cubiella, Joaquín Diez, Maria Soledad Bujanda, Luis Lanas, Angel Jover, Rodrigo Hernández, Vicent Quintero, Enrique Herreros-Villanueva, Marta Martín, Ana Carmen Pérez-Palacios, Rosa Arroyo, Rocio Castells, Antoni Gironella, Meritxell |
author_sort | Duran-Sanchon, Saray |
collection | PubMed |
description | Background: Circulating microRNA (miRNA) analysis is a growing research field. However, it usually requires an endogenous control or housekeeping (HK) in order to normalize expression of specific miRNAs throughout different samples. Unfortunately, no adequate HK for circulating miRNA analysis is still known in the colorectal cancer (CRC) context whereas several have been suggested. Hence, our aims were to validate the previously suggested miR-1228-3p as HK for CRC studies, to compare its suitability with the widely used miR-16-5p, and to evaluate the influence of hemolysis on both miRNAs. Methods: We analyzed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) the expression of miR-1228-3p, miR-16-5p and the spike-in cel-miR-39 in a set of 297 plasmas (92 CRC, 101 advanced adenomas -AA-, and 100 controls) and 213 serum samples (59 CRC, 74 AA and 80 controls). We also analyzed both miRNAs depending on the hemolysis degree in 7 plasmas and 31 serums. Results: Levels of miR-1228-3p and miR-16-5p did not show significant differences between groups although miR-16-5p exhibited more variability in plasma and serum samples. Importantly, the combination of cel-miR-39 and miR-1228-3p was the most stable one. Moreover, we observed that miR-16-5p was significantly influenced by hemolysis in contrast with miR-1228-3p that exhibited no correlation with this confounding factor in both biofluids. Conclusion: MiR-1228-3p has been validated as an adequate endogenous control for circulating miRNA analysis in CRC and AA liquid biopsies. |
format | Online Article Text |
id | pubmed-7022916 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-70229162020-03-12 Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients Duran-Sanchon, Saray Vila-Navarro, Elena Marcuello, Maria Lozano, Juan José Muñoz, Jenifer Cubiella, Joaquín Diez, Maria Soledad Bujanda, Luis Lanas, Angel Jover, Rodrigo Hernández, Vicent Quintero, Enrique Herreros-Villanueva, Marta Martín, Ana Carmen Pérez-Palacios, Rosa Arroyo, Rocio Castells, Antoni Gironella, Meritxell Biomolecules Article Background: Circulating microRNA (miRNA) analysis is a growing research field. However, it usually requires an endogenous control or housekeeping (HK) in order to normalize expression of specific miRNAs throughout different samples. Unfortunately, no adequate HK for circulating miRNA analysis is still known in the colorectal cancer (CRC) context whereas several have been suggested. Hence, our aims were to validate the previously suggested miR-1228-3p as HK for CRC studies, to compare its suitability with the widely used miR-16-5p, and to evaluate the influence of hemolysis on both miRNAs. Methods: We analyzed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) the expression of miR-1228-3p, miR-16-5p and the spike-in cel-miR-39 in a set of 297 plasmas (92 CRC, 101 advanced adenomas -AA-, and 100 controls) and 213 serum samples (59 CRC, 74 AA and 80 controls). We also analyzed both miRNAs depending on the hemolysis degree in 7 plasmas and 31 serums. Results: Levels of miR-1228-3p and miR-16-5p did not show significant differences between groups although miR-16-5p exhibited more variability in plasma and serum samples. Importantly, the combination of cel-miR-39 and miR-1228-3p was the most stable one. Moreover, we observed that miR-16-5p was significantly influenced by hemolysis in contrast with miR-1228-3p that exhibited no correlation with this confounding factor in both biofluids. Conclusion: MiR-1228-3p has been validated as an adequate endogenous control for circulating miRNA analysis in CRC and AA liquid biopsies. MDPI 2019-12-20 /pmc/articles/PMC7022916/ /pubmed/31877644 http://dx.doi.org/10.3390/biom10010016 Text en © 2019 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 Duran-Sanchon, Saray Vila-Navarro, Elena Marcuello, Maria Lozano, Juan José Muñoz, Jenifer Cubiella, Joaquín Diez, Maria Soledad Bujanda, Luis Lanas, Angel Jover, Rodrigo Hernández, Vicent Quintero, Enrique Herreros-Villanueva, Marta Martín, Ana Carmen Pérez-Palacios, Rosa Arroyo, Rocio Castells, Antoni Gironella, Meritxell Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients |
title | Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients |
title_full | Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients |
title_fullStr | Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients |
title_full_unstemmed | Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients |
title_short | Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients |
title_sort | validation of mir-1228-3p as housekeeping for microrna analysis in liquid biopsies from colorectal cancer patients |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7022916/ https://www.ncbi.nlm.nih.gov/pubmed/31877644 http://dx.doi.org/10.3390/biom10010016 |
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