title: Detection of microRNAs in archival cytology urine smears.
abstract: 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.
authors.affiliation: Department of Medicine (DIMED), Surgical Pathology & Cytopathology Unit, University of Padua, Padua, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Simonato
authors.forename: Laura
authors.initials: L
authors.lastname: Ventura
authors.forename: Nicola
authors.initials: N
authors.lastname: Sartori
authors.forename: Rocco
authors.initials: R
authors.lastname: Cappellesso
authors.forename: Matteo
authors.initials: M
authors.lastname: Fassan
authors.forename: Lill-Tove
authors.initials: LT
authors.lastname: Busund
authors.forename: Ambrogio
authors.initials: A
authors.lastname: Fassina
mesh_terms: D005240:Feasibility Studies; D006801:Humans; D035683:MicroRNAs / Q000652:urine*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D035683:MicroRNAs