Prostate cancer (PCa) represents the most frequently diagnosed cancer in men. Cisplatin, also known as cis-diamminedichloroplatinum (DDP), is a standard chemotherapeutic agent used to treat PCa, and DDP resistance remains one important obstacle in DDP-based chemotherapy. In our research, we found miR-425-5p was down-regulated in PCa and even lower in DDP-resistant PCa determined by quantitative polymerase chain reaction; in contrast, GSK3β mRNA expression was upregulated in PCa and even higher in DDP-resistant PCa. Moreover, there was a modest but significant inverse correlation between the expression of GSK3β mRNA and miR-425-5p. Functional experiments showed that miR-425-5p mimic inhibited DDP resistance as evidenced by a promoted apoptosis rate (flow cytometry) and suppressed cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay) and expressions of MDR1 andMRP1 (western blotting) in DU145/DDP and PC3/DDP cells. Luciferase reporter assay and RNA immunoprecipitation identifiedGSK3β was a potential target of miR-425-5p. The effect ofmiR-425-5pmimic on DDP resistance was partially reversed by pcDNA-GSK3β. Mechanically, miR-425-5p mimic reduced expression of β-catenin, cyclin D1 and C-myc, which was further blocked when GSK3β overexpressed. In vivo experiments, recovery of GSK3β prevented xenograft tumor growth and DDP resistance in the presence of miR-425-5p mimic. To sum up, miR-425-5p upregulation might sensitize human PCa to DDP by targeting GSK3β and inactivating the Wnt/β-catenin signaling pathway.