Mori Folium ethanol extracts induce ferroptosis and suppress gastric cancer progression by inhibiting the AKT/GSK3β/NRF2 axis.

BACKGROUND: Mori Folium, the leaf of Morus alba L., is a traditional Chinese medicine (TCM) known for its diverse pharmacological activities, including anti-inflammatory and immunomodulatory effects. While the Morus alba itself has been reported to contain various bioactive compounds with anticancer properties, the anticancer activity of Mori Folium and its underlying mechanisms remain insufficiently understood.

PURPOSE: This study aimed to investigate the effects of Mori Folium ethanol extracts (MFEE) on gastric cancer (GC) and to elucidate its underlying mechanisms.

METHODS: To investigate the anti-GC properties of MFEE, CCK-8, colony formation, EdU, flow cytometry, and soft agar, scratch, and transwell assays were employed. Western blot was employed to analyze the expression of ferroptotic proteins, while ferroptotic cellular events were also assessed, including iron accumulation, GSH levels, reactive oxygen species (ROS) production, mitochondrial changes, and lipid peroxidation. The chemical profile of MFEE was characterized using a UPLC-ESI-MS/MS system. Additionally, network pharmacology analysis was performed to investigate the potential anti-GC mechanisms of MFEE. Finally, the in vivo anti-cancer effects of MFEE were evaluated using a subcutaneous mouse model, with hematoxylin and eosin (H&E) and immunohistochemistry (IHC) staining to assess histopathological and molecular changes.

RESULTS: This study demonstrated that MFEE suppresses GC cell proliferation, blocks the G1-S cell cycle transition, and inhibits migration and invasion by promoting Fe²⁺ accumulation, increasing MDA levels and ROS, depleting GSH, and downregulating the expression of xCT and GPX4, thereby inducing ferroptosis. Chemical analysis identified 1596 phytochemicals, including 35 bioactive compounds. The induction of ferroptosis by MFEE was associated with the inhibition of the PI3K/AKT signaling pathway, modulating the AKT/GSK3β/NRF2 axis. Activation of AKT by SC79 was found to mitigate MF-induced ferroptosis. Notably, MFEE enhanced the chemosensitivity of GC cells to cisplatin, potentially through ferroptosis induction.

CONCLUSION: This study revealed that MFEE induces ferroptosis in GC cells by modulating the PI3K/AKT signaling pathway, enhancing chemosensitivity to cisplatin, and providing a potential therapeutic strategy for GC.