Fig. 8: Clinical relevance of the ACTL6A-GCLC-GSH metabolism axis in ferroptosis of GC cells.

a Immunoblotting analysis of ACTL6A and GCLC protein levels in PDX tumors of 4 cases. b. The mice were treated with BSO (750 mg/kg/day) for two weeks. c–f Tumor volumes of PDX tumors, including case1 (c), case2 (d), case3 (e), and case4 (f). The data are presented as the means ± SD, n = 6 for each group in case 1 and 3, n = 4 for each group in case 2 and 4. g–j Representative fluorescent images of PDX frozen sections stained with DCFH-DA (g) and C11-BODIPY (h). Scale bars represent 200 μm. Relative ROS levels (i) and lipid peroxidation levels (j) were counted by image J and presented as bar graphs. The data are presented as the means ± SD, n = 5 for each group. BODIPY GFP represents lipid peroxidation; BODIPY tomato represents the original color of BODIPY. k Schematic metabolic map of [U-¹⁵N]-labeled GSH de novo synthesis. Gln glutamine, Glu glutamate, Cys cysteine, Gly glycine, γ-GC γ-glutamyl-cysteine, GSH glutathione. l–n. Relative intracellular pool levels of glutamine, glutamate, and GSH in case1 (ACTL6A high) PDX tumors (l). Incorporation of nitrogen atoms from [U-¹⁵N] glutamine into glutamine (gln), glutamate (glu), and GSH (m). Relative GSH/Gln ratios (n). Data are presented as the means ± SD, n = 3 for each group. o Kaplan–Meier curves of the overall survival of GC patients with different expression levels of ACTL6A and GCLC, and log-rank analysis was used to test for significance. p Representative images showing the correlation of ACTL6A and GCLC staining in human GC tissue microarray samples. Scale bars represent 50 μm. P values were determined by two-way ANOVA followed by Tukey test for panels c–f, unpaired two-tailed T test for panels i, j, l–n, h, and the log-rank test for panel o.