Reference miRNAs for colorectal cancer: analysis and verification of current data

Abstract MicroRNAs (miRNAs) hold great promise in cancer research. The use of appropriate reference miRNAs for normalization of qPCR data is crucial for accurate expression analysis. We present here analysis and verification of current data, proposing a workflow strategy for identification of refere...

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Autores principales:	E. Danese, A. M Minicozzi, M. Benati, E. Paviati, G. Lima-Oliveira, M. Gusella, F. Pasini, G. L Salvagno, M. Montagnana, G. Lippi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/19c153b502724e3cbcaba4cdd7c0f836
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spelling	oai:doaj.org-article:19c153b502724e3cbcaba4cdd7c0f8362021-12-02T16:06:33ZReference miRNAs for colorectal cancer: analysis and verification of current data10.1038/s41598-017-08784-32045-2322https://doaj.org/article/19c153b502724e3cbcaba4cdd7c0f8362017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08784-3https://doaj.org/toc/2045-2322Abstract MicroRNAs (miRNAs) hold great promise in cancer research. The use of appropriate reference miRNAs for normalization of qPCR data is crucial for accurate expression analysis. We present here analysis and verification of current data, proposing a workflow strategy for identification of reference miRNAs in colorectal cancer (CRC). We performed a systematic review of studies aimed to identify stable reference miRNAs in CRC through high-throughput screening. Among the candidate miRNAs selected from the literature we excluded those predicted to target oncogenes or tumor suppressor gene. We then assessed the expression levels of the remaining candidates in exosomes, plasma and tissue samples from CRC patients and healthy controls. The expression stability was evaluated by box-plot, ∆Cq analysis, NormFinder and BestKeeper statistical algorithms. The effects of normalisers on the relative quantification of the oncogenic miR-1290 was also assessed. Our results consistently showed that different combinations of miR-520d, miR-1228 and miR-345 provided the most stably expressed reference miRNAs in the three biological matrices. We identified suitable reference miRNAs for future miRNA expression studies in exosomes plasma and tissues CRC samples. We also provided a novel conceptual framework that overcome the need of performing ex novo identification of suitable reference genes in single experimental systems.E. DaneseA. M MinicozziM. BenatiE. PaviatiG. Lima-OliveiraM. GusellaF. PasiniG. L SalvagnoM. MontagnanaG. LippiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q E. Danese A. M Minicozzi M. Benati E. Paviati G. Lima-Oliveira M. Gusella F. Pasini G. L Salvagno M. Montagnana G. Lippi Reference miRNAs for colorectal cancer: analysis and verification of current data
description	Abstract MicroRNAs (miRNAs) hold great promise in cancer research. The use of appropriate reference miRNAs for normalization of qPCR data is crucial for accurate expression analysis. We present here analysis and verification of current data, proposing a workflow strategy for identification of reference miRNAs in colorectal cancer (CRC). We performed a systematic review of studies aimed to identify stable reference miRNAs in CRC through high-throughput screening. Among the candidate miRNAs selected from the literature we excluded those predicted to target oncogenes or tumor suppressor gene. We then assessed the expression levels of the remaining candidates in exosomes, plasma and tissue samples from CRC patients and healthy controls. The expression stability was evaluated by box-plot, ∆Cq analysis, NormFinder and BestKeeper statistical algorithms. The effects of normalisers on the relative quantification of the oncogenic miR-1290 was also assessed. Our results consistently showed that different combinations of miR-520d, miR-1228 and miR-345 provided the most stably expressed reference miRNAs in the three biological matrices. We identified suitable reference miRNAs for future miRNA expression studies in exosomes plasma and tissues CRC samples. We also provided a novel conceptual framework that overcome the need of performing ex novo identification of suitable reference genes in single experimental systems.
format	article
author	E. Danese A. M Minicozzi M. Benati E. Paviati G. Lima-Oliveira M. Gusella F. Pasini G. L Salvagno M. Montagnana G. Lippi
author_facet	E. Danese A. M Minicozzi M. Benati E. Paviati G. Lima-Oliveira M. Gusella F. Pasini G. L Salvagno M. Montagnana G. Lippi
author_sort	E. Danese
title	Reference miRNAs for colorectal cancer: analysis and verification of current data
title_short	Reference miRNAs for colorectal cancer: analysis and verification of current data
title_full	Reference miRNAs for colorectal cancer: analysis and verification of current data
title_fullStr	Reference miRNAs for colorectal cancer: analysis and verification of current data
title_full_unstemmed	Reference miRNAs for colorectal cancer: analysis and verification of current data
title_sort	reference mirnas for colorectal cancer: analysis and verification of current data
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/19c153b502724e3cbcaba4cdd7c0f836
work_keys_str_mv	AT edanese referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT amminicozzi referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT mbenati referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT epaviati referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT glimaoliveira referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT mgusella referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT fpasini referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT glsalvagno referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT mmontagnana referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata AT glippi referencemirnasforcolorectalcanceranalysisandverificationofcurrentdata
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Reference miRNAs for colorectal cancer: analysis and verification of current data

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