Acetylshikonin induces ferroptosis via the lipid peroxidation pathway in osteosarcoma cells.

Osteosarcoma, a prevalent primary malignant bone tumor, primarily affects adolescents and young adults. Current treatment strategies involve a combination of surgical intervention and chemotherapy. However, the effectiveness of chemotherapy is constrained by considerable challenges, such as drug resistance and insensitivity. Ferroptosis, a form of programmed cell death that is distinct from apoptosis, presents a promising alternative target for cancer therapy. Ferroptosis is characterized by iron‑dependent lipid peroxidation, producing reactive oxygen species (ROS) and suppressing glutathione peroxidase 4 (GPX4). Notably, ferroptosis circumvents the conventional mechanisms associated with apoptosis. Inducing ferroptosis in cancer cells may help overcome drug resistance and enhance the effectiveness of existing treatments, including chemotherapy, radiotherapy and immunotherapy. Acetylshikonin is a derivative of naphthoquinone that possesses anticancer properties. However, the effects of acetylshikonin on the treatment of osteosarcoma and the mechanisms by which it induces cancer cell death remain unclear. The present study aimed to investigate the potential of acetylshikonin to induce apoptosis in osteosarcoma cells. Using cell viability assays, ROS detection, mitochondrial membrane potential analysis and ferroptosis inhibitor rescue experiments, the results demonstrated that acetylshikonin significantly reduced the viability of osteosarcoma cell lines while exhibiting low toxicity to normal cells. Mechanistically, acetylshikonin induced the production of ROS, disrupted the mitochondrial membrane potential and promoted lipid peroxidation, ultimately leading to ferroptosis. Additionally, treatment with acetylshikonin led to decreased levels of GPX4 and increased intracellular ferrous ion (Fe²+) concentrations, further supporting its role in the induction of ferroptosis. In conclusion, the current study emphasized the potential of acetylshikonin as an effective agent in inducing ferroptosis in osteosarcoma cells. Acetylshikonin reduced osteosarcoma cell viability and selectively promoted ferroptosis by increasing ROS production, disrupting mitochondrial function and enhancing lipid peroxidation. Furthermore, its ability to downregulate GPX4 and increase intracellular Fe2+ levels indicated its role in triggering ferroptosis. These findings suggest that acetylshikonin may be a valuable therapeutic candidate for the treatment of osteosarcoma, potentially improving outcomes and addressing the limitations of current therapies.