Figure 4. Levobupivacaine up-regulates and activates p53 in NSCLC cells. (A and B) The A549 and A427 cells were treated with levobupivacaine (2 mM) or equal volume saline. The total expression and nucleus accumulation of p53 was measured by Western blot analysis in the cells. (C) The A549 and A427 cells were treated with p53 overexpression vectors. (D and E) The A549 and A427 cells were co-treated with 5 mmol/L erastin or ferrostatin (1 mmol/L) and p53 overexpression vectors. The cell growth was analyzed by MTT assays. (F–J) The A549 and A427 cells were treated with p53 overexpression vectors or control vectors. (F) The levels of ROS were measure by flow cytometry analysis in the cells. (G and H) The levels of iron and Fe2+ were analyzed by Iron Assay Kit. (I and J) The expression of GPX4, SLC7A11, and β-actin was measured by Western blot analysis in the cells. Data are presented as mean ± SD. Statistic significant differences were indicated: *P < 0.05, **P < 0.01, ***P < 0.001.