Aim: Radical prostatectomy (RP) is the most frequent frontline PCa treatment. Biochemical recurrence (BCR) after radical prostatectomy occurs in 20%–40% of patients, but only 30% of these patients demonstrate cancer progression. Sensitive and specific markers of RP effectiveness are needed. Cell-free miRNAs from blood plasma packed in extracellular vesicles (EVs), namely the expression of 14 miRNAs before and one week after RP, were studied in comparison with their expression in EVs of benign prostatic hyperplasia patients and healthy donors in the present manuscript. Materials and Methods: Plasma EVs isolation was performed using an aggregation-precipitation protocol. MiRNA was isolated using the Guanidine isothiocyanate/Octanoic Acid Protocol. MiRNAs expression was assessed by reverse transcription and quantitative RT-PCR. Results: It was shown that 11 of the 72 studied miRNA ratios changed significantly after RP. Moreover, one of two miRNAs (miR-125b and miR-30e) took part in each miRNA ratio whose relative expression changed after RP. Conclusion: RP causes differential expression of plasma EVs miRNA. The obtained results indicate the prominent role of miR-125b and miR-30e in response to radical therapy. The study of miRNA expression in dynamics and in different biofluid fractions is required to assess the potential of extracellular miRNAs as sensitive biomarkers of therapy and to select their optimal source.