Everolimus, a known inhibitor of the
mammalian target of rapamycin (mTOR), has shown uncertain efficacy in treating
hepatoblastoma. This study delves into the potential anti-
hepatoblastoma properties of
everolimus and its intricate relationship with autophagy and ferroptosis, both in vitro and in vivo. In vivo,
tumor tissue from
hepatoblastoma patient and human
hepatoblastoma cell line HuH-6 were xenografted into nude mice to establish xenograft models for observing the effect of
everolimus on
tumor growth. In vitro, HuH-6 cells were cultured to evaluate the anti-
hepatoblastoma activity of
everolimus. Transmission electron microscopy and
microtubule-associated proteins 1 light chain 3 (LC3),
beclin 1, and p62
protein expressions were employed to investigate autophagy. Additionally, indicators of cell apoptosis,
reactive oxygen species (ROS) and
proteins associated with ferroptosis were measured to evaluate ferroptosis. The results demonstrate that
everolimus treatment effectively induced the formation of autophagosomes in
hepatoblastoma cells, upregulated the LC3II/I ratio and
beclin 1 expression, and downregulated p62 expression, indicating an enhanced autophagy level both in vitro and in vivo. Furthermore,
everolimus treatment induced cell apoptosis, increased ROS level, elevated concentrations of
malondialdehyde,
4-hydroxynonenal, and
iron content, while reducing the ratio of
glutathione/oxidized glutathione, and downregulating the
protein expression of
glutathione peroxidase 4 and solute carrier family 7 member 11, suggesting its ability to induce ferroptosis in
hepatoblastoma cells. Importantly, the induction of ferroptosis by
everolimus was significantly reversed in the presence of
autophinib, an autophagy inhibitor, indicating the autophagy-dependent of
everolimus-induced ferroptosis. Taken together, these findings suggest that
everolimus holds promise as an effective anti-
hepatoblastoma drug, with its mechanism of action potentially involving the induction of autophagy-dependent ferroptosis in
hepatoblastoma cells.