Fig. 3
Metabolic adaptations in drug-resistant HCC cells. a Nontargeted metabolomics profiling of parental and Huh-7 LR cells. b FFA profiling in parental and Huh-7 LR cells. c Enrichment analysis of upregulated metabolites (FC > 1.2, p < 0.05) in Huh-7 LR cells compared with parental cells. d Seahorse extracellular flux analysis of glycolytic activity (ECAR) in parental and drug-resistant cells. Testing method: Unpaired Student’s t-test. e Visualization of intracellular lipid droplet accumulation during drug resistance development. Lipid droplets (green), the cell membrane (red), and the nucleus (blue). Scale bar = 25 μm. IC50 values: Huh-7 LR_lenvatinib (S0: 13.02 μM; S1: 19.15 μM; S2: 36.40 μM; S3: 50.61 μM); Huh-7 SR_sorafenib (S0: 6.77 μM; S1: 9.46 μM; S2: 13.53 μM; S3: 19.46 μM). f Correlation analysis between the intracellular lipid droplet content and drug IC50 in drug-resistant cells. Testing method: Pearson’s correlation coefficient test. g Integrated metabolomic (four replicates per cell type, averaged in pairs), transcriptomic (two replicates per cell type), and supplementary kit-based assay profiling to map metabolic adaptations in drug-resistant cells. h High-resolution 13C-glucose metabolic flux analysis of glycolysis, the TCA cycle, and glutathione metabolism pathways. The “M+number” indicates the number of additional ¹³C atoms in the metabolite molecule. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001
