Fig. 5

Modulation of energy generation and metabolism in HK1⁻HK2⁺ Hep3B/shHK2^DOX cells in response to FDG/DPI/PER treatment. a FDG/DPI/PER combination treatment acutely decreases the OCR of Hep3B cells. FDG (250 μM), DPI (15 nM), and/or PER (5 μM) were added to cultured Hep3B cells, as single agents or in the combinations shown, at time 0. The OCR was measured by the Seahorse assay. Oligomycin (2 μM), FCCP (0.1 μM), and Antimycin (2 μM) were used to indicate the fraction of respiration coupled to oxidative ATP production, maximum mitochondrial respiration, and non-mitochondrial respiration, respectively. b OCR in Hep3B cells after 3, 8, and 24 h of drug exposure; concentrations are as in panel a. c The HK2i/DPI/PER triple combination decreases liver cancer cellular energy levels. After an 8-h drug treatment, AMP, ADP, ATP, creatine, and P-creatine amounts in Hep3B cells were determined by LC-MS. d Changes in pool sizes of glycolysis and TCA cycle metabolites, as well as purine and pyrimidine nucleotides, after vehicle, FDG/DPI, or FDG/DPI/PER treatment in Hep3B cells. F1,6BP fructose 1,6-biphosphate, G3P glyceraldehyde 3-phosphate, DHAP dihydroxyacetone phosphate, 3PG 3-phospho-glycerate, PEP phosphoenolpyruvate, Cit citrate, α-KG α-ketoglutarate, Succ succinate, Fum fumarate, Mal malate. Each data point in a and b represents mean ± SEM of five samples, and each data point in c and d represents mean ± SD of triplicate samples. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001