The influence of maximal androgen blockade and radical prostatectomy on urinary extracellular vesicle miRNA expression.

Prostate cancer (PC) is one of the most common oncological diseases. Maximal androgen blockade (MAB) is used at different stages of treatment, including prior to radical prostatectomy (RP). The data on the expression of extracellular miRNAs involved in carcinogenesis before and after MAB/RP's are demanded for the selection of miRNAs markers for the diagnostics of PC and the effectiveness of PC treatment as well as insight of these miRNAs and their regulated genes involvement in oncogenesis. The aim of this study was to screen 13 miRNAs associated with PC from urine extracellular vesicles (EVs) of healthy donors (HD) and PC patients before and after MAB, RP, and MAB followed RP in looking for correlations with PC and PC treatment/clinical characteristics followed by identification of the most distinctive miRNA signatures as set of markers for the future extensive verification. Urine samples were collected from HD and PC patients, both those who had undergone neoadjuvant therapy and those who had not, prior to radical prostatectomy (RP) and one-week post-surgery. EVs were isolated using the aggregation-precipitation method, and miRNAs within EVs were extracted using glass fiber sorbents. The quantification of the 13 miRNAs was performed using reverse transcription TaqMan PCR (RT-PCR), with subsequent paired ratio-based normalization followed by the selection of miRNA ratios differentially expressed between the comparison groups from the set of all miRNA pairs. A comparative analysis of the relative expression of miRNAs in the urine of PC patients versus HD was conducted, revealing 49 ratios of differentially expressed miRNAs influenced by therapy. The study identified specific miRNAs with significant expression changes due to various treatments: influence of MAB, RP, and RP in patients after neoadjuvant MAB, as well as under the influence of MAB followed by RP. The relative expression of the extracellular miRNAs studied was influenced by the type and combination of various PC treatments, which indicates the prospects of specific miRNA predictors of PC treatment effectiveness.