• Sanguinarine (San) significantly inhibits HCC cell proliferation, migration, and EMT while promoting apoptosis in vitro and suppresses tumor growth in vivo.
• San induces cuproptosis by upregulating FDX1, LIAS, HSP70, and lipoylated DLAT aggregation, increasing copper levels, and reducing mitochondrial membrane potential and glutathione/pyruvate levels.
• San synergizes with copper ionophores (Elesclomol-CuCl2) to enhance cuproptosis, and FDX1 silencing attenuates its anti-HCC effects.
• Molecular docking, SPR, and CETSA confirm that San directly binds to FDX1, enhancing its thermostability, thereby targeting the FDX1/LIAS/DLAT/HSP70 axis.