Urinary exosome microRNA signatures as a non-invasive prognostic biomarker for prostate cancer

We aimed to identify urinary exosomal miRNAs associated with PCa metastasis and develop a non-invasive risk-scoring model for PCa metastasis in this study. MiRNA profiles were examined using the Taqman low-density miRNA array (TLDA). Megaplex reverse transcription reactions and pre-amplification reactions were performed to increase the quantity of cDNA for miRNA expression analysis using the Megaplex PreAmp Primers Human Pool A and TaqMan PreAmp Master Mix (Thermo Fisher Scientific). MiRNA expression was evaluated via the TLDA panel A v2.0 (Thermo Fisher Scientific). Raw data were processed using the QuantStudio Real-Time PCR Software (Thermo Fisher Scientific) to determine a cycle threshold (Ct) value for each miRNA. We analyzed miRNA expression profiles of 42 patients (19 localized and 23 metastatic) using the Taqman low-density miRNA array (TLDA) containing 381 miRNAs. Exosomes were isolated from 5 mL of urine, and Total RNA was isolated from the pellets using TRIzol.