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Introduction:
MicroRNAs (miRs) are small, non-coding segments of regulatory RNA that are powerful biomarkers of disease severity and prognosis. Previous work from this group indicated the potential for identification of miRNAs that play a role in urothelial carcinoma of the bladder (UCB). In this study we isolated RNA from cell-free urine to identify a miRNA profile that could be used as a non-invasive diagnostic assay to detect the presence of UCB and provide a discriminatory signature for different stages of progression.
Materials & Methods: Total RNA was isolated from cell-free urine of patients with UCB and controls. Samples were grouped according to grade and stage. MiRNAs were profiled by qRT-PCR array on pooled samples within each group. Validation of miRNAs was performed on individual samples using qRT-PCR.
236 miRNAs were detected in at least one pooled sample; the number of miRNAs detected correlated with disease progression. The control group and the >=T2 group expressed 8 and 228 miRNAs, respectively. Of miRNAs present in both cancer and non-cancer groups, 13 had significantly higher levels in the cancer group. Statistical analysis adjusted for multiple comparisons demonstrated differences between groups based on miRNA expression levels including a panel of miRNAs that discriminated between cancer and cancer-free patients with high sensitivity and specificity.
Conclusions: We demonstrated successful isolation of miRNAs from cell-free urine. Utilizing this non-invasive assay, we identified miRNAs capable of discriminating between cancer-free patients and patients with UCB, providing evidence that miRNA profiling holds promise for the development of valuable clinical tools.