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Detection of MicroRNAs in Archival Cytology Urine Smears

MicroRNAs’ dysregulation and profiling have been demonstrated to be clinically relevant in urothelial carcinoma (UC). Urine cytology is commonly used as the mainstay non-invasive test for secondary prevention and follow-up of UC patients. Ancillary tools are needed to support cytopathologists in the...

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Autores principales: Simonato, Francesca, Ventura, Laura, Sartori, Nicola, Cappellesso, Rocco, Fassan, Matteo, Busund, Lill-Tove, Fassina, Ambrogio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3585351/
https://www.ncbi.nlm.nih.gov/pubmed/23469001
http://dx.doi.org/10.1371/journal.pone.0057490
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author Simonato, Francesca
Ventura, Laura
Sartori, Nicola
Cappellesso, Rocco
Fassan, Matteo
Busund, Lill-Tove
Fassina, Ambrogio
author_facet Simonato, Francesca
Ventura, Laura
Sartori, Nicola
Cappellesso, Rocco
Fassan, Matteo
Busund, Lill-Tove
Fassina, Ambrogio
author_sort Simonato, Francesca
collection PubMed
description MicroRNAs’ dysregulation and profiling have been demonstrated to be clinically relevant in urothelial carcinoma (UC). Urine cytology is commonly used as the mainstay non-invasive test for secondary prevention and follow-up of UC patients. Ancillary tools are needed to support cytopathologists in the diagnosis of low-grade UC. The feasibility and reliability of microRNAs profiling by qRT-PCR analysis (miR-145 and miR-205) in archival routine urine cytology smears (affected by fixation/staining [Papanicolau] and room temperature storage) was tested in a series of 15 non-neoplastic and 10 UC urine specimens. Only samples with >5,000 urothelial cells and with <50% of inflammatory cells/red blood cells clusters were considered. Overall, a satisfactory amount of total RNA was obtained from all the considered samples (mean 1.27±1.43 µg, range 0.06–4.60 µg). Twenty nanograms of total RNA have been calculated to be the minimal total RNA concentration for reliable and reproducible miRNAs expression profiling analysis of archival cytological smears (slope = -3.4084; R-squared = 0.99; efficiency = 1.94). miR-145 and miR-205 were significantly downregulated in UC samples in comparison to non-tumor controls. These findings demonstrate that urine archival cytology smears are suitable for obtaining high-quality RNA to be used in microRNAs expression profiling. Further studies should investigate if miRNAs profiling can be successfully translated into clinical practice as diagnostic or prognostic markers.
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spelling pubmed-35853512013-03-06 Detection of MicroRNAs in Archival Cytology Urine Smears Simonato, Francesca Ventura, Laura Sartori, Nicola Cappellesso, Rocco Fassan, Matteo Busund, Lill-Tove Fassina, Ambrogio PLoS One Research Article MicroRNAs’ dysregulation and profiling have been demonstrated to be clinically relevant in urothelial carcinoma (UC). Urine cytology is commonly used as the mainstay non-invasive test for secondary prevention and follow-up of UC patients. Ancillary tools are needed to support cytopathologists in the diagnosis of low-grade UC. The feasibility and reliability of microRNAs profiling by qRT-PCR analysis (miR-145 and miR-205) in archival routine urine cytology smears (affected by fixation/staining [Papanicolau] and room temperature storage) was tested in a series of 15 non-neoplastic and 10 UC urine specimens. Only samples with >5,000 urothelial cells and with <50% of inflammatory cells/red blood cells clusters were considered. Overall, a satisfactory amount of total RNA was obtained from all the considered samples (mean 1.27±1.43 µg, range 0.06–4.60 µg). Twenty nanograms of total RNA have been calculated to be the minimal total RNA concentration for reliable and reproducible miRNAs expression profiling analysis of archival cytological smears (slope = -3.4084; R-squared = 0.99; efficiency = 1.94). miR-145 and miR-205 were significantly downregulated in UC samples in comparison to non-tumor controls. These findings demonstrate that urine archival cytology smears are suitable for obtaining high-quality RNA to be used in microRNAs expression profiling. Further studies should investigate if miRNAs profiling can be successfully translated into clinical practice as diagnostic or prognostic markers. Public Library of Science 2013-02-28 /pmc/articles/PMC3585351/ /pubmed/23469001 http://dx.doi.org/10.1371/journal.pone.0057490 Text en © 2013 Simonato et al http://creativecommons.org/licenses/by/4.0/ 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 author and source are properly credited.
spellingShingle Research Article
Simonato, Francesca
Ventura, Laura
Sartori, Nicola
Cappellesso, Rocco
Fassan, Matteo
Busund, Lill-Tove
Fassina, Ambrogio
Detection of MicroRNAs in Archival Cytology Urine Smears
title Detection of MicroRNAs in Archival Cytology Urine Smears
title_full Detection of MicroRNAs in Archival Cytology Urine Smears
title_fullStr Detection of MicroRNAs in Archival Cytology Urine Smears
title_full_unstemmed Detection of MicroRNAs in Archival Cytology Urine Smears
title_short Detection of MicroRNAs in Archival Cytology Urine Smears
title_sort detection of micrornas in archival cytology urine smears
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3585351/
https://www.ncbi.nlm.nih.gov/pubmed/23469001
http://dx.doi.org/10.1371/journal.pone.0057490
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