Purpose. To determine whether Ski‐interacting protein (SKIP) regulates TGF‐β1‐stimulated expression of urokinase‐type plasminogen activator (uPA), matrix metalloproteinase‐9 (MMP‐9), and uPA Inhibitor (PAI‐1) in the androgen‐independent human prostate cancer cell model. Materials and Methods. PC‐3 prostate cancer cell line was used. The role of SKIP was evaluated using synthetic small interference RNA (siRNA) compounds. The expression of uPA, MMP‐9, and PAI‐1 was evaluated by zymography assays, RT‐PCR, and promoter transactivation analysis. Results. In PC‐3 cells TGF‐β1 treatment stimulated uPA, PAI‐1, and MMP‐9 expressions. The knockdown of SKIP in PC‐3 cells enhanced the basal level of uPA, and TGF‐β1 treatment inhibited uPA production. Both PAI‐1 and MMP‐9 production levels were increased in response to TGF‐β1. The ectopic expression of SKIP inhibited both TGF‐β1‐induced uPA and MMP‐9 promoter transactivation, while PAI‐1 promoter response to the factor was unaffected. Conclusions. SKIP regulates the expression of uPA, PAI‐1, and MMP‐9 stimulated by TGF‐β1 in PC‐3 cells. Thus, SKIP is implicated in the regulation of extracellular matrix degradation and can therefore be suggested as a novel therapeutic target in prostate cancer treatment.