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 i...

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Main Authors: Francesca Simonato, Laura Ventura, Nicola Sartori, Rocco Cappellesso, Matteo Fassan, Lill-Tove Busund, Ambrogio Fassina
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2013-01-01
Series:PLoS ONE
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23469001/pdf/?tool=EBI
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author Francesca Simonato
Laura Ventura
Nicola Sartori
Rocco Cappellesso
Matteo Fassan
Lill-Tove Busund
Ambrogio Fassina
author_facet Francesca Simonato
Laura Ventura
Nicola Sartori
Rocco Cappellesso
Matteo Fassan
Lill-Tove Busund
Ambrogio Fassina
author_sort Francesca Simonato
collection DOAJ
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 doaj.art-1dc93f8f621b4592a76e8a12886c186e2022-12-21T23:41:05ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5749010.1371/journal.pone.0057490Detection of microRNAs in archival cytology urine smears.Francesca SimonatoLaura VenturaNicola SartoriRocco CappellessoMatteo FassanLill-Tove BusundAmbrogio FassinaMicroRNAs' 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.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23469001/pdf/?tool=EBI
spellingShingle Francesca Simonato
Laura Ventura
Nicola Sartori
Rocco Cappellesso
Matteo Fassan
Lill-Tove Busund
Ambrogio Fassina
Detection of microRNAs in archival cytology urine smears.
PLoS ONE
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
url https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23469001/pdf/?tool=EBI
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