fig6

Figure 6. Fra-1 inhibits ferroptosis and induces chemoresistance in GC cells by activating the PPP metabolic pathway. (A) Detection of ROS levels in GC cells AGS after overexpression of Fra-1, and overexpression of Fra-1 along with treatment with 6AN (4 μM), using a ROS detection kit; (B-G) Detection of the effect of Fra-1 on GSH, MDA, Fe²⁺ levels in GC cells AGS and HGC27 using GSH, MDA, Fe²⁺ ELISA assay kit after overexpression/silencing of Fra-1; (H-K) RT-qPCR assay to detect mRNA expression levels of ferroptosis negatively related factors GPX4, SLCA7A11, and FTH1 in GC cells AGS and HGC27 after overexpression/silencing of Fra-1; (L) Detection of protein expression levels of ferroptosis negatively related factors GPX4 and SLCA7A11 in GC cells AGS and HGC27 after overexpression/silencing of Fra-1 using western blot assay; (M-R) Detection of the effect of Fra-1 on GSH, MDA, Fe²⁺ content in GC cells AGS and HGC27 after overexpression of Fra-1, and overexpression of Fra-1 along with treatment with 6AN (4 μM), using GSH, MDA, Fe²⁺ ELISA assay kit; (S-V) Detection of cell proliferation ability in GC cells AGS and HGC27 after overexpression/silencing of Fra-1, overexpression/silencing of Fra-1 along with the addition of ferroptosis inducer erastin (8 μM), and treatment with CDDP (10 μM) for 24 h using the EDU Cell Proliferation Detection Kit. All experiments were performed with three technical replicates. ns, no significant difference; ^*P < 0.05; ^**P < 0.01; ^***P < 0.001. Fra-1: Fos-related antigen-1; GC: gastric cancer; PPP: pentose phosphate pathway; ROS: reactive oxygen species; GSH: glutathione; MDA: malondialdehyde; CDDP: cisplatin; EDU: 5-ethynyl-2’-deoxyuridine.