Fig. 2: KEAP1WT-dependent SAT1 upregulation enhances the synergy between polyamines and KRAS inhibitors.
A Relative viabilities of MIAPACA2 with KEAP1 knockout (KO) and H23 cells with KEAP1 overexpression (OE) treated with gradient concentrations of spermine and sotorasib or adagrasib for 72 h. 2D surface response for cell viability and 3D surface Bliss synergy response were shown. (n = 3 independent experiments). B Schematic depiction of polyamine metabolism. Created in BioRender. Bian, Y. (2025) https://BioRender.com/3p0ref1. C The volcano plots showed differentially expressed genes (DEGs) in MIAPACA2 and H23 cells treated with KRASG12C inhibitors (sotorasib: MIAPACA2, IC50 = 40 nM; H23, IC50 = 500 nM; adagrasib: MIAPACA2, IC50 = 60 nM; H23, IC50 = 100 nM, 72 h) by RNA-seq analysis. ODC1, SAT1, SMOX, and PAOX are critical in polyamine metabolism. “DN” means downregulated in the KRAS inhibitors treatment group, and “UP” means upregulated in the KRAS inhibitors treatment group (|log2FC | > 0.4 and p < 0.05). D qRT-PCR and WB verified the mRNA and protein expression level of ODC1, SAT1, SMOX, and PAOX in MIAPACA2 and H23 cell lines treated with sotorasib or adagraisb (IC50) for 72 h. (n = 3 independent experiments) Data were analyzed by two-tailed Student’s t-test and were presented by mean ± SD. Source data are provided as a Source Data file. E Polyamine metabolites level in MIAPACA2 and H23 cells following KRAS inhibitors treatment (IC50, 72 h). (n = 3 independent experiments) Data were analyzed by two-tailed Student’s t-test and were presented by mean ± SD. Source data are provided as a Source Data file. F Polyamine metabolites level in MIAPACA2 (NC and SAT1-KO) and H23 (NC and SAT1-OE) cells. (n = 3 independent experiments) Data were analyzed by two-tailed Student’s t-test and were presented by mean ± SD. Source data are provided as a Source Data file. G Relative viabilities of MIAPACA2 cells (NC and SAT1-KO group) and H23 cells (NC and SAT1-OE group) treated with spermine (100 nM) or spermidine (100 nM), KRAS inhibitors (IC50), or combination for 72 h. (n = 3 independent experiments) Data were analyzed by two-tailed Student’s t-test and were presented by mean ± SD. Source data are provided as a Source Data file. H qRT-PCR and WB showed the expression level of SAT1 in MIAPACA2 cells (NC, KEAP1-KO, KEAP1-WT, and KEAP1-MU group) and H23 cells (NC and KEAP1-OE group). (n = 3 independent experiments) Data were analyzed by two-tailed Student’s t-test and were presented by mean ± SD. Source data are provided as a Source Data file. Source data are provided as a Source Data file.
