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Detection of microRNAs in archival cytology urine smears.

Simonato F, Ventura L, Sartori N, Cappellesso R, Fassan M, Busund LT, Fassina A - PLoS ONE (2013)

Bottom Line: 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).Further studies should investigate if miRNAs profiling can be successfully translated into clinical practice as diagnostic or prognostic markers.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine (DIMED), Surgical Pathology & Cytopathology Unit, University of Padua, Padua, Italy.

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.

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Related in: MedlinePlus

Selection of the cytology specimens.Only cases with >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were selected. Cases largely constituted by red blood cells or inflammatory cells were excluded.
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pone-0057490-g001: Selection of the cytology specimens.Only cases with >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were selected. Cases largely constituted by red blood cells or inflammatory cells were excluded.

Mentions: The presence of >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were appointed as minimum accepted cellularity. In neoplastic specimens, only slides with a proportion of tumor cells >25% were selected. Slides with >50% of urothelial cells obscured by either white or red blood cells were excluded (Figure 1).


Detection of microRNAs in archival cytology urine smears.

Simonato F, Ventura L, Sartori N, Cappellesso R, Fassan M, Busund LT, Fassina A - PLoS ONE (2013)

Selection of the cytology specimens.Only cases with >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were selected. Cases largely constituted by red blood cells or inflammatory cells were excluded.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585351&req=5

pone-0057490-g001: Selection of the cytology specimens.Only cases with >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were selected. Cases largely constituted by red blood cells or inflammatory cells were excluded.
Mentions: The presence of >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were appointed as minimum accepted cellularity. In neoplastic specimens, only slides with a proportion of tumor cells >25% were selected. Slides with >50% of urothelial cells obscured by either white or red blood cells were excluded (Figure 1).

Bottom Line: 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).Further studies should investigate if miRNAs profiling can be successfully translated into clinical practice as diagnostic or prognostic markers.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine (DIMED), Surgical Pathology & Cytopathology Unit, University of Padua, Padua, Italy.

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.

Show MeSH
Related in: MedlinePlus