Fig. 4

Inhibition of fatty acid oxidation sensitizes HK1⁻HK2⁺ liver cancer cells to the cytotoxicity of the HK2 inhibition/DPI combination. a FDG/DPI or FDG/ROT combinations induce ACC phosphorylation. HK1⁻HK2⁺ liver cancer cells were treated with the indicated drug(s) for 24 h prior to analysis of ACC phosphorylation. FDG, 1 mM. DPI, 100 nM. ROT, 100 nM. b Clinical liver cancer samples show upregulated expression of genes involved in fatty acid elongation and downregulated expression of genes involved in FAO compared to normal liver tissues. Data were obtained from Oncomine (http://www.oncomine.com). c PER sensitizes Hep3B cells to FDG/DPI treatment. Cells were treated with FDG, plus or minus 5 μM PER, for 72 h and then assayed for viable cells by the MTT assay. Open surface, [DPI + FDG + vehicle] treatment. Filled surface, [DPI + FDG + PER] treatment. d PER sensitizes HK1⁻HK2⁺ liver cancer cells to the cytotoxicity of FDG/DPI treatment. Triplicate wells of (left) Hep3B or (right) Huh7 cells were treated with FDG (250 μM), DPI (15 nM), and/or PER (5 μM) for 72 h, followed by trypan blue staining to determine cell viability. Viability % = 100% × (trypan blue negative cells/total cells). Data are means ± SD. e PER rescues FDG/DPI-induced lipid droplet depletion in Hep3B cells. Hep3B cells were treated with DMSO, FDG (1 mM)/DPI (100 nM), or FDG (1 mM)/DPI (100 nM)/PER (5 μM) for 24 h, followed by Oil Red staining of intracellular lipid droplets. Representative images with indicated treatments are shown. Scale bars, 30 μM. f FDG, DPI, and/or PER modulation of AMPKα activation, mTOR pathway inactivation, and apoptosis induction in Hep3B/shHK2^DOX cells. Cells were treated with FDG (250 μM), DPI (15 nM), and/or PER (5 μM) for 24 h, and cell extracts were examined by Western blotting