Figure 2 from Oncogenic Mutant <i>p53</i> Sensitizes Non–Small Cell Lung Cancer Cells to Proteasome Inhibition via Oxidative Stress–Dependent Induction of Mitochondrial Apoptosis

<p><i>Onc-p53</i> NSCLC cells exhibit basal and PI-induced oxidative stress. <b>A,</b> Average oxidized (GSSG) vs. reduced (GSH) levels of GSH obtained from metabolomic profiles (DepMap) of NSCLC cell lines that either express Onc-p53 protein (Onc-p53) or lack p53 protein expression due to homozygous deletion/frameshift/premature termination mutations (loss of function; LOF). Data were analyzed using the unpaired <i>t</i> test. <b>B,</b> GSH concentration was determined in lysates of the indicated cell lines. <b>C,</b> Relative GSH concentrations were determined in lysates of H1975-Control vs. H1975-p53KO cells. <b>D,</b> Levels of ROS were determined in the indicated cell lines using a fluorescent ROS detection assay and plotted as mean fluorescence units (MFU). <b>E,</b> H460 or H1975 cells were treated with vehicle, BTZ (5 nmol/L), NAC (1 mmol/L), or BTZ + NAC for 24 hours. Relative ROS levels normalized to vehicle treatment for each cell line were determined by staining with DCFH-DA followed by flow cytometry. <b>F,</b> H1975 cells were treated as per <b>E</b> for 48 hours, and cell viability was assessed by trypan blue exclusion. All pairwise comparisons were made using the Student <i>t</i> test. *, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ***, <i>P</i> < 0.005. Error bars indicate ± 1.0 SD.</p>

Duke Scholars

Author Eziafa I Oduah Medicine, Medical Oncology