Targeting ferroptosis in rhabdomyosarcoma cells

Area of Science:

• Oncology
• Cell Biology
• Biochemistry

Background:

• Rhabdomyosarcoma (RMS) cells may be susceptible to oxidative stress.
• Erastin, a glutamate/cystine antiporter inhibitor, induces cell death by increasing reactive oxygen species (ROS) via glutathione (GSH) depletion.
• Understanding ferroptosis mechanisms in RMS is crucial for developing novel therapies.

Purpose of the Study:

• To investigate the susceptibility of RMS cells to Erastin-induced ferroptosis.
• To elucidate the molecular mechanisms, including the roles of protein kinase C (PKC) and NADPH oxidase (NOX) pathways, in Erastin-induced cell death in RMS.

Main Methods:

• RMS cell lines were treated with Erastin.
• Assessed GSH depletion, ROS production, and lipid peroxidation.
• Utilized pharmacological inhibitors (Ferrostatin-1, Liproxstatin-1, α-Tocopherol, GSH, Deferoxamine, Bisindolylmaleimide I, Gö6976, Diphenyleneiodonium, GKT137831).
• Performed genetic knockdown of PKCα.

Main Results:

• Erastin induced GSH depletion, ROS production, and lipid peroxidation in RMS cells.
• Inhibitors of lipid peroxidation, ROS scavengers, and an iron chelator demonstrated ferroptosis.
• PKC inhibitors (broad-spectrum and PKCα/β-selective) and PKCα knockdown significantly reduced Erastin-induced cell death.
• NOX inhibitors (broad-spectrum and NOX1/4-selective) decreased Erastin-stimulated ROS, lipid ROS, and cell death.

Conclusions:

• RMS cells are sensitive to Erastin-induced ferroptosis.
• PKCα and NOX pathways are critical mediators of ferroptosis in RMS.
• These findings support the development of redox-based therapeutic strategies for rhabdomyosarcoma.