Metformin and Ferroptosis in Lung Cancer

A549 and H1299 cells (Cell Bank of the Typical Culture Preservation Committee, Chinese Academy of Sciences) were cultured in RPMI-1640 medium (Gibco, China ) supplemented with 10% fetal bovine serum (FBS) (Biological Industries, Israel) and 100 µg/mL PenStrep. The cells cultured at 37 °C in 5% CO₂ humidified chamber were passaged when they grew to 80% confluence. Cells were inoculated into six-well plates with 1 × 10⁶ cells per well and cultured under normal conditions for 48 h. Subsequently, the cells were treated with 0.5 mM, 1.0 mM or 2.0 mM metformin [24 (link)] (Met, S1950, Selleck, China) for 48 h with or without the supplement of 10 µM Ferrostatin-1 (Fer-1, S7243, Selleck, China).