Lycorine suppresses DPP3-mediated Nrf2 signaling, thereby reducing SLC7A11 levels to trigger ferroptosis in colorectal cancer cells

BACKGROUND: Colorectal cancer (CRC) exhibits high morbidity/mortality with poor late-stage survival, creating an urgent need for effective therapies. Although lycorine, a pyrrolophenanthridine alkaloid isolated from the traditional Chinese herb Lycoris radiata, has been confirmed to exhibit significant anti-CRC activity. the exact biological mechanism of its action remains unclear PURPOSE: The present study aimed to clarify whether lycorine exerts its anti-CRC effect by inducing ferroptosis in CRC cells (HCT-116 and RKO), and to elucidate its specific molecular mechanism. METHODS: Human CRC cell lines HCT-116 and RKO were used in vitro: lentiviral transfection was applied to establish cell models with DPP3/solute carrier family 7 member 11 (SLC7A11) overexpression or knockdown, and cells were treated with lycorine, nuclear factor erythroid 2-related factor 2 (Nrf2) activator dimethyl fumarate (DMF), or Nrf2 inhibitor Nrf2 inhibitor (ML385), followed by detection of cell proliferation, apoptosis, invasion, reactive oxygen species (ROS), intracellular Fe²⁺ level, GSH/GSSG ratio, Malondialdehyde (MDA)level, 4-Hydroxynonenal (4-HNE) level and expression of related molecules. The ferroptosis-dependent effect was verified using inhibitors such as Deferoxamine(DFO). Molecular docking and enzyme activity experiments were conducted to elucidate the interaction between lycorine and DPP3. For in vivo validation, CRC xenograft models were constructed in nude mice, with tumor volume/weight monitored and hematoxylin and eosin (HE) staining, Ki67 Antigen (Ki67) detection, MDA detection, 4-HNE detection and molecular analyses of tumor tissues performed; RESULTS: Our results demonstrate that lycorine significantly inhibits cell proliferation, invasion, and migration, while promoting apoptosis and ROS generation, indicating its effective suppression of the malignant phenotype in CRC. Further studies reveal that lycorine induces ferroptosis in CRC cells. However, overexpression of DPP3 or SLC7A11, or activation of Nrf2, attenuates the anticancer activity of lycorine. Knockdown of DPP3 suppresses the malignant phenotype of CRC cells, an effect that can be significantly reversed by Nrf2 activator treatment or SLC7A11 overexpression. Additionally, SLC7A11 knockdown markedly counteracts the tumor-promoting effects induced by DPP3 overexpression. Molecular docking and enzymatic activity assays confirm the direct binding of lycorine to DPP3. Ferroptosis inhibitors significantly alleviate lycorine-induced cell death. In vivo experiments further validate this regulatory mechanism. CONCLUSION: Lycorine can directly bind to and inhibit DPP3, thereby blocking the Nrf2 signaling pathway and downregulating SLC7A11. This disruption of GSH metabolism leads to the accumulation of ROS and iron overload, ultimately inducing ferroptosis in CRC cells.