Xanthohumol Triggers Pyroptotic in Prostate Cancer Cells via the Caspase-3/GSDME Signaling Pathway

Xanthohumol (XN), a naturally occurring flavonoid characterized by the presence of prenyl moieties and obtained from hop plants (<i>Humulus lupulus</i> L.), has garnered growing interest in the scientific community owing to its diverse biological activities, including anti-inflammatory, anticancer, and antioxidant effects. However, its antitumor mechanisms, especially the inhibitory impact and related molecular pathways in prostate cancer, are not yet fully elucidated. This study investigated the effects of XN on prostate cancer and explored its underlying molecular mechanisms. The antiproliferative effect of XN on prostate cancer cells was assessed using the sulforhodamine B assay. Cellular morphological changes were examined by microscopy. Pyroptosis induction following XN treatment was evaluated via flow cytometry and Western blot analysis. Following treatment with XN, prostate cancer cells exhibited characteristic morphological changes consistent with pyroptosis. Protein analysis revealed that XN triggers pyroptosis primarily via the caspase-3/GSDME. The attenuation of XN-induced, GSDME-dependent pyroptosis by the caspase-3-specific inhibitor <i>Z-DEVD-fmk</i> further supported this mechanism. Furthermore, our results indicate that XN promotes the accumulation of reactive oxygen species (ROS) and reduces mitochondrial membrane potential, thereby activating the mitochondrial intrinsic pathway and leading to cytochrome c release, which subsequently triggers caspase-3 activation and the cleavage of GSDME, and ultimately induces pyroptosis. XN induced pyroptosis in prostate cancer cells through the mitochondrial intrinsic pathway, offering novel strategic insights for the treatment of prostate cancer and the development of innovative therapeutic agents.