MicroRNAs in urine supernatant as potential non-invasive markers for bladder cancer detection

Abstract

Urinary bladder carcinoma contributes to 4% of newly diagnosed oncological diseases in the Czech Republic. Biomarkers for its early non-invasive detection are therefore highly desirable. Urine seems to be an ideal source of such biomarkers due to the content of cell-free nucleic acids, especially microRNAs (miRNAs).To find potential biomarkers among miRNAs in urine supernatant, we examined in total 109 individuals (36 controls and 73 bladder cancer patients) in three phases. In the first - discovery - phase, microarray cards with 381 miRNAs were used for miRNA analysis of 13 controls and 46 bladder cancer patients. In the second - verification - phase, the results of this first phase were verified on the same groups of subjects by single-target qPCR assays for the selected miRNAs. For the third - validation - phase, new independent samples of urine supernatant (23 controls and 27 bladder cancer patients) were analyzed using single-target qPCR assays for 13 verified in the previous phase. The results of all phases were normalized to miR-191, miR-28-3p, and miR-200b, which were selected as suitable for our study by the qBase+®.We found that miR-125b, miR-30b, miR-204, miR-99a, and miR-532-3p are significantly down-regulated in patients' urine supernatant. In our experiments, the analysis of miR-125 levels provided the highest AUC (0.801) with 95.65% specificity and 59.26% sensitivity, the analysis of miR-99a lead to AUC (0.738) with 82.61% specificity and 74.07% sensitivity. We demonstrate that levels of these miRNAs could potentially serve as promising diagnostic markers for the non-invasive diagnostics of bladder cancer.

Genome‐wide identification of urinary cell‐free microRNAs for non‐invasive detection of bladder cancer

“…In addition to BCA detection, our DxScore enabled also to distinguish between NMIBC and MIBC and low‐grade and high‐grade form of NMIBC and significantly decreased in the urinary samples collected after surgery in disease‐free patients compared to pre‐operative samples. When compared to previous studies , our study is performed on the larger cohort of well‐defined patients with BCA and healthy controls, showing more promising analytical characteristics of cell‐free urinary miRNAs in BCA than described before.…”

Pathological significance and prognostic implications of heme oxygenase 1 expression in non-muscle-invasive bladder cancer: Correlation with cell proliferation, angiogenesis, lymphangiogenesis and expression of VEGFs and COX-2

“…In addition, various molecular and biological markers, such as survivin, interleukins, microRNAs, have also been reviewed (4,5). However, there are no clinically definitive markers for NMIBC, as tumor growth, invasion, metastasis and recurrence are regulated by complex underlying mechanisms.…”

Urinary expression of let-7c cluster as non-invasive tool to assess the risk of disease progression in patients with high grade non-muscle invasive bladder Cancer: a pilot study

“…In line with let-7c tumorsuppressor role, our analysis of the cluster expression levels in HG-NMIBC patient's tissue showed that let-7c, as well as miR-99a and miR-125b, were highly downregulated compared to normal tissue samples. In urine, let-7c, as well as the other let-7c cluster members, miR-99a and miR-125b has been found down-regulated in BC versus HC [11,22,23]. In the current study, we investigated the potential value of urinary let-7c cluster as noninvasive diagnostic/prognostic biomarker of HG-NMIBC patients.…”