Background and Purpose: Aberrant overexpression of UHRF1 has been reported in several cancer types and UHRF1 is regarded as a novel drug target for cancer therapy. However, no UHRF1-targeted specific small compound inhibitor has been registered in clinical trials. Experimental Approach: Network pharmacology together with molecular docking were used to screened a natural molecule bank for PCa treatment. The expression of related protein or mRNA were evaluated by WB and RT-PCR. The ubiquitination levels were assessed by WB. CCK8 assess was used to measuring cells viability. Additionally, PCa cells cycle were analysed by cytofluorimetry, genomic DNA methylation was assessed by Dot blot analysis. Cellular senescence was assessed by Senescence-Associated β-Galactosidase Staining Kit. DU145 cell xenograft models were used to assess the in vivo effect of DSG inhibition. Key Results: Identified DSG as a new natural compound specifically targeting UHRF1 protein degradation through the ubiquitin-proteasome pathway, DSG-induced UHRF1 protein degradation reduced the level of genomic DNA methylation, and re-activated the expression of such TSGs as p21, p16 and LXN, thereby resulted in cell cycle arrest, cell senescence and reduced DU145 xenograft tumor growth. Altogether, clarified DSG anticancer mechanism as an epigenetic regulatory drug for the treatment of PCa. Conclusions and Implications: Our results first time identified DSG which extract from natural plants specifically targeting UHRF1 protein. This vpresent study provided a promising strategy to discover new molecule-targeted drug from natural compounds. KEYWORDS: Traditional Chinese Medicine; Prostate cancer; Diosgenin; DNA methylation; Tumor suppressor genes;