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.
Highlights
Bladder cancer represents the fourth male most common cancer in the western world and urothelial cell carcinoma (UC; previously designated as transitional carcinoma [TCC]) accounts for approximately 95% of bladder malignancies and is the most common tumor detected by urine cytology [1]
Two distinct phenotypic and molecular pathways have been demonstrated in urothelial carcinoma (UC): i) the superficial papillary carcinoma, accounting for more than 80% of bladder tumors, with tendency to recur locally, with rare invasion and metastasis; ii) and the invasive non-papillary bladder tumor, without known papillary precursor, which is commonly associated with carcinoma in situ (CIS) with unfavourable prognosis [2,3]
Urine cytology is widely and routinely employed as: i) the preliminary analysis in the evaluation of patients presenting with haematuria or painful urination suggestive for urinary system pathology [4]; ii) screening test for the early detection of bladder cancer in selected populations exposed to known bladder carcinogens; and iii) the mainstay in the follow-up of patients with a history of malignancy involving the urinary tract [4,5]
Summary
Bladder cancer represents the fourth male most common cancer in the western world and urothelial cell carcinoma (UC; previously designated as transitional carcinoma [TCC]) accounts for approximately 95% of bladder malignancies and is the most common tumor detected by urine cytology [1]. MiRNA profiling methods have been showed to be standardizable and of clinical impact in human cancers [19,20]. Such characteristics pinpointed miRNAs as suitable biomarkers to be detected even in scant degraded cytology samples affected by fixation/staining and room temperature storage such as routine diagnostics urine smears
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