Background: Prostate cancer (PCa) is a common malignancy in men characterized by high heterogeneity and frequent therapeutic resistance, particularly in advanced stages. Ginsenoside Rh2 (Rh2), a bioactive compound extracted from red ginseng, has demonstrated antitumor activity in various cancers, but its role and mechanisms in PCa remain unclear. Methods: We systematically investigated the effects of Rh2 on DU145 PCa cells through functional assays and transcriptome sequencing. Cell viability, proliferation, migration, invasion, and apoptosis were evaluated. RNA Sequencing and downstream analyses were performed to identify potential regulatory targets and pathways. MAFF knockdown, overexpression, and combined MAPK inhibition experiments to validate its functional relevance. Results: Rh2 treatment significantly suppressed DU145 cell proliferation, migration, and invasion while promoting apoptosis. Transcriptomic profiling revealed pronounced downregulation of the transcription factor MAFF. Western blot analysis confirmed that Rh2 reduced MAFF expression and inhibited phosphorylation of p38, ERK, and JNK. Silencing MAFF recapitulated the tumor-suppressive effects of Rh2, whereas MAFF overexpression enhanced malignant phenotypes, including proliferation, motility, and resistance to apoptosis. Importantly, the p38 MAPK inhibitor BIRB796 partially reversed the oncogenic effects driven by MAFF overexpression, underscoring a MAFF-MAPK regulatory axis in PCa. Conclusion: Rh2 may inhibit malignant phenotypes of PCa cells in vitro by downregulating MAFF and attenuating MAPK pathway activation. The identification of the Rh2-MAFF-MAPK axis provides novel mechanistic insight into Rh2's anticancer effects and highlights MAFF as a potential molecular target for PCa.

prostate cancer, ginsenoside rh2, rna-seq, MAFF, MAPK