Astragaloside IV is associated with TFRC-dependent ferroptosis and JAK2/STAT3 pathway inhibition in osteosarcoma...

Osteosarcoma is a common malignant bone tumor in children and adolescents. Conventional chemotherapy has drug resistance and side effects, and new treatments are needed. Astragaloside IV (AS-IV) has anti-tumor effects, but its mechanism in osteosarcoma is unclear.

MG63/Saos2 osteosarcoma cells were divided into control, AS-IV (20/40 μM), Erastin (ferroptosis inducer), AS-IV + Lip-1 (ferroptosis inhibitor), AS-IV + short hairpin RNA negative control (shNC), and AS-IV + short hairpin RNA Transferrin Receptor 1 (shTFRC) (transfection of shTFRC lentiviral vector) groups. Cell counting kit-8, flow cytometry, immunofluorescence, kit detection, quantitative real-time polymerase chain reaction, and Western blot were used to detect cell viability, apoptosis, reactive oxygen species (ROS), iron metabolism, oxidative stress, and Janus Kinase 2/Signal Transducers and Activators of Transcription 3 (JAK2/STAT3) pathway-related indicators.

AS-IV could concentration-dependently inhibit the viability of MG63 and Saos2 cells, promote cell apoptosis and ROS production, upregulate the levels of Fe²⁺, labile iron pool (LIP) and malondialdehyde (MDA), downregulate the expression of Ferritin Heavy Chain 1 (FTH1) and Glutathione Peroxidase 4 (GPX4) mRNA and the activities of superoxide dismutase (SOD) and catalase (CAT), and significantly inhibit the phosphorylation of JAK2 and STAT3. Lip-1 could reverse these effects. Moreover, after TFRC knockdown, the regulatory effects of AS-IV on the viability, apoptosis, iron metabolism, oxidative stress and JAK2/STAT3 pathway of osteosarcoma cells were significantly weakened.

AS-IV is associated with ferroptosis induction and JAK2/STAT3 pathway inhibition in osteosarcoma cells, and these effects are attenuated by TFRC knockdown, providing new targets and theoretical basis for the treatment of osteosarcoma.