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Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR

BACKGROUND: MicroRNAs have recently taken centre stage as short non-coding RNAs that regulate mRNA expression. AIM/METHODS: To assess the feasibility of using microRNA techniques on routinely processed tissues, the accessibility of two representative microRNAs was examined by real-time quantitative...

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Autores principales: Siebolts, U, Varnholt, H, Drebber, U, Dienes, H-P, Wickenhauser, C, Odenthal, M
Formato: Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2603282/
https://www.ncbi.nlm.nih.gov/pubmed/18755714
http://dx.doi.org/10.1136/jcp.2008.058339
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author Siebolts, U
Varnholt, H
Drebber, U
Dienes, H-P
Wickenhauser, C
Odenthal, M
author_facet Siebolts, U
Varnholt, H
Drebber, U
Dienes, H-P
Wickenhauser, C
Odenthal, M
author_sort Siebolts, U
collection PubMed
description BACKGROUND: MicroRNAs have recently taken centre stage as short non-coding RNAs that regulate mRNA expression. AIM/METHODS: To assess the feasibility of using microRNA techniques on routinely processed tissues, the accessibility of two representative microRNAs was examined by real-time quantitative PCR in 86 human formalin-fixed paraffin-embedded (FFPE) samples from liver, breast, bone marrow, lymphatic tissues and colon. Murine liver was used to analyse the influence of fixation time and different fixatives. RESULTS: High-quality microRNA was successfully extracted from routinely processed formalin-fixed tissues, resembling PCR amplification results from snap-frozen material analysed in parallel. While fixation time did not affect microRNA accessibility, non-buffered formalin or fixative supplements such as glutaraldehyde influenced PCR results. Storage of human tissues for up to 7 years did not cause a significant deterioration of microRNA. However, microRNA quality in human archival material following routine processing 10–20 years ago was decreased. Oxidation by ambient air during storage and fixation in non-buffered formalin is a possible reason for loss of microRNA quality. CONCLUSION: The assessment of microRNAs in readily obtained formalin-fixed paraffin-embedded samples is a highly promising tool in molecular pathology when similarly treated samples are analysed. Therefore, microRNA analyses will gain wider acceptance as an adjunct to morphological tissue assessment in routine pathology and retrospective studies.
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spelling pubmed-26032822009-01-01 Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR Siebolts, U Varnholt, H Drebber, U Dienes, H-P Wickenhauser, C Odenthal, M J Clin Pathol Original Articles BACKGROUND: MicroRNAs have recently taken centre stage as short non-coding RNAs that regulate mRNA expression. AIM/METHODS: To assess the feasibility of using microRNA techniques on routinely processed tissues, the accessibility of two representative microRNAs was examined by real-time quantitative PCR in 86 human formalin-fixed paraffin-embedded (FFPE) samples from liver, breast, bone marrow, lymphatic tissues and colon. Murine liver was used to analyse the influence of fixation time and different fixatives. RESULTS: High-quality microRNA was successfully extracted from routinely processed formalin-fixed tissues, resembling PCR amplification results from snap-frozen material analysed in parallel. While fixation time did not affect microRNA accessibility, non-buffered formalin or fixative supplements such as glutaraldehyde influenced PCR results. Storage of human tissues for up to 7 years did not cause a significant deterioration of microRNA. However, microRNA quality in human archival material following routine processing 10–20 years ago was decreased. Oxidation by ambient air during storage and fixation in non-buffered formalin is a possible reason for loss of microRNA quality. CONCLUSION: The assessment of microRNAs in readily obtained formalin-fixed paraffin-embedded samples is a highly promising tool in molecular pathology when similarly treated samples are analysed. Therefore, microRNA analyses will gain wider acceptance as an adjunct to morphological tissue assessment in routine pathology and retrospective studies. BMJ Publishing Group 2009-01 2008-08-28 /pmc/articles/PMC2603282/ /pubmed/18755714 http://dx.doi.org/10.1136/jcp.2008.058339 Text en © Siebolts et al 2009 http://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Siebolts, U
Varnholt, H
Drebber, U
Dienes, H-P
Wickenhauser, C
Odenthal, M
Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR
title Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR
title_full Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR
title_fullStr Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR
title_full_unstemmed Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR
title_short Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR
title_sort tissues from routine pathology archives are suitable for microrna analyses by quantitative pcr
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2603282/
https://www.ncbi.nlm.nih.gov/pubmed/18755714
http://dx.doi.org/10.1136/jcp.2008.058339
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