Detection of MicroRNAs in Archival Cytology Urine Smears
Permanent link
https://hdl.handle.net/10037/6030Date
2013Type
Journal articleTidsskriftartikkel
Peer reviewed
Author
Simonato, Francesca; Ventura, Laura; Sartori, Nicola; Cappellesso, Rocco; Fassan, Matteo; Busund, Lill-Tove; Fassina, AmbrogioAbstract
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 1.2761.43 mg,
range 0.06–4.60 mg). 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; Rsquared
= 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.
Publisher
Public Library of Science (PLoS)Citation
PLoS ONE (2013), vol. 8(2): e57490Metadata
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