Fig. 7

The shHK2/DPI/PER combination does not inhibit proliferation or tumor progression for HK1⁺HK2⁺ cancer cells but is effective in inhibiting proliferation and progression for the isogenic HK1^KOHK2⁺ cancer cells. a Effect of the shHK2/DPI/PER combination on isogenic HK1⁺HK2⁺ H460WT/shHK2^DOX and H460HK1^KO/shHK2^DOX cell proliferation. Cells were pretreated with vehicle or DOX for 2 days, followed by a 72-h treatment with DPI in the indicated concentration range, in the presence or absence of DOX and the presence or absence of PER (5 μM). Upper panels, MTT assay values were normalized to control samples (-DOX, -DPI, -PER) of the individual isogenic cell lines. Data are expressed as means ± SD. Lower panels, representative images of MTT assay results. b The shHK2/DPI/PER combination triggers apoptosis in H460HK1^KO/shHK2^DOX cells but not in H460WT/shHK2^DOX cells. Cells were pretreated with vehicle or DOX for 2 days, followed by a 24-h treatment with 15 nM DPI, 5 μM PER, and DOX, both as single agents and in combination (Comb). Cell lysates were examined by Western blot for HK1, HK2, and cleaved caspase 3 expression. c The shHK2/DPI/PER combination inhibits the progression of H460HK1^KO/shHK2^DOX tumor xenografts but not H460WT/shHK2^DOX tumor xenografts. When tumors reached 200 mm³ (day 0), xenografts were randomized into five groups (n = 5 per group) receiving vehicle, DOX in the diet, DPI (2 mg/kg, daily i.p.), and PER (30 mg/kg, daily i.p.), both as single agents and in combination. Data are expressed as means ± SEM. **P < 0.01. ***P < 0.001. NS not significant. d The shHK2/DPI/PER combination triggers apoptosis in H460HK1^KO/shHK2^DOX tumor xenografts but not H460WT/shHK2^DOX tumor xenografts. Three representative xenograft tumors in each group in panel c were homogenized for Western blotting analyses of indicated proteins