Direct quantitative detection for cell-free miR-155 in urine: a potential role in diagnosis and prognosis for non-muscle invasive bladder cancer

Abstract

High recurrence rates of non-muscle invasive bladder cancer (NMIBC) in patients require lifelong testing and monitoring. The aim of this study is to develop a simplified RT-qPCR method (RT-qPCR-D) which directly quantifies cell-free miR-155 in urine without RNA extraction, and assess it as a potential tool in NMIBC detection. A pilot study including 60 urine samples was used to investigate the feasibility of RT-qPCR-D in detecting cell-free miR-155. Then, miR-155 levels were quantified in a large independent cohort of urine from 162 NIMBC patients, 76 cystitis patients, and 86 healthy donors using the RT-qPCR-D method. Changes of cell-free miR-155 before and after operation were also analyzed in 32 NIMBC patients. In pilot study, we found a significant linear association between RT-qPCR and RT-qPCR-D in urinary miR-155 detection. Both methods showed cell-free miR-155 were significantly increased in NMIBC patients, and could reflect their expression in tissues. Then, the increased expression of cell-free miR-155 was successfully validated in 162 NIMBC patients when compared with cystitis patients and healthy donors. Moreover, it distinguished NMIBC patients from others with 80.2% sensitivity and 84.6% specificity, which was superior to urine cytology. Cell-free miR-155 correlated with NMIBC stage and grade, and was an independent factor for predicting recurrence and progression to muscle invasion. In addition, cell-free miR-155 was significantly decreased after NMIBC patients underwent transurethral bladder resection. In conclusion, detection of cell-free miR-155 in urine using RT-qPCR-D is a simple and noninvasive approach which may be used for NMIBC diagnosis and prognosis prediction.

Biomarkers for Bladder Cancer Diagnosis and Surveillance: A Comprehensive Review

“…Xan et al described that high levels of miR-155 were associated with lower recurrence-free and PFS. miR-155 expression also distinguished the cluster of patients with BC from the cluster without BC with a sensitivity of 80.2% and a specificity of 84.6% [85]. Evaluation of specific methylation pattern in miR-137, miR-124-2, miR-124-3, and miR-9-in voided urine samples of BC patients with NMIBC presented, for recurrence detection, a 61.5% sensitivity and 74.0% specificity.…”

The Diagnostic and Prognostic Significance of MicroRNA-21 in Non-muscle Invasive Bladder Tumors

“…In recent years, studies have been published on the prognostic significance of different miRNA molecules in patients with BC. In 2016, Zhang et al (48) reported that miRNA-155 analysis in cell-free urine samples of NMIBC patients had diagnostic value with 85.8% sensitivity. In another study investigating the role of miRNA-203 in predicting treatment response in patients planned to start cisplatin-based chemotherapy for BC, it was reported that low miRNA-203 level predicted progression and poor prognosis while overexpression of miRNA-203 may increase the sensitivity to cisplatin by directly stimulating apoptosis (49).…”

Secreted miR-210-3p as non-invasive biomarker in clear cell renal cell carcinoma

“…The identification of molecular markers in body fluids (e.g., sera and urine), which can be used as noninvasive diagnostic, prognostic and surveillance markers in ccRCC management, is one of the most ambitious challenges in oncologic research [20]. Of note, also miRNAs are acquiring a great potential as novel cancer biomarkers in urological carcinomas [16,[21][22][23][24][25]. To date, the increased interest on non-invasive biomarkers, allowed by use of novel methodologies (such as next generation sequencing, single-cell sequencing approaches and digital PCR) has greatly improved the translational potential of miRNAs into clinical research [16].…”