Sanguinarine chloride inducing ferroptosis and downregulating GPX4 expression in liver cancer: An integrated in silico, in vitro and in vivo study

Abstract

Ferroptosis, an iron-dependent form of regulated cell death driven by excessive lipid peroxidation, has been increasingly implicated in the development and progression of liver cancer. Despite sanguinarine chloride (SC) exhibiting antitumor activity in a number of cancer models, whether it modulates ferroptosis in liver cancer remains unclear. The present study systematically investigated the ferroptosis-inducing effects of SC and the underlying molecular mechanisms in liver cancer. Molecular docking and molecular dynamics simulations suggested that SC interacts with glutathione peroxidase 4 (GPX4) with favorable affinity and forms a stable complex. Functional assays showed that SC significantly inhibited proliferation and reduced the viability of HepG2 and Huh7 cells. Mechanistically, SC markedly decreased GPX4 protein expression, leading to the accumulation of lipid reactive oxygen species and malondialdehyde, together with characteristic mitochondrial changes associated with ferroptosis, including increased membrane density and loss of cristae. SC-induced cell death was partially rescued by the ferroptosis inhibitors ferrostatin-1 and deferoxamine, further supporting the involvement of ferroptosis. Consistently, in a xenograft model, SC suppressed GPX4 expression, promoted ferroptosis and significantly inhibited tumor growth. Collectively, these findings indicate that SC exerts antitumor effects in liver cancer, at least in part, by inducing ferroptosis through GPX4 downregulation, supporting further investigation of SC as a potential therapeutic candidate.