Figure 6.

Metformin (Met) continued to exert its anticancer effects on cell proliferation, cell cycle and apoptosis in QGP-1-R and H727-R cells. (A,B) To measure cell proliferation, everolimus-resistant QGP-1-R and H727R cell were treated with metformin (1–10 mM) and everolimus (Eve 10–100 nM) or their combination for 24 h and then with BrdU for 2 h. Experiments were repeated 3 times and each determination was done in triplicate. Values represent mean (± SD.) * = p < 0.05, *** = p < 0.001, **** = p < 0.0001 vs. corresponding basal. (C,D) QGP-1-R and H727-R cells were treated with metformin (10 mM) and everolimus (10 nM) or their combination for 24 h and their cell cycle was analyzed by flow cytometry. Experiments were repeated at least 3 times. Values represent mean (± SD.) * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 vs. corresponding basal. Statistical analysis was performed with a one-way ANOVA test followed by Dunnett’s post-hoc test. (E,F) The graph showed the percentage of apoptotic cells after metformin (10 mM) and everolimus (10 nM) treatment, alone or in combination, compared to basal in QGP-1-R and H727-R cells. Only metformin 10 mM significantly promoted apoptosis. Values represent mean ± SD of 3 experiments. ** = p < 0.01, *** = p < 0.001 vs. corresponding basal. Statistical analysis was performed with a one-way ANOVA test followed by Dunnett’s post-hoc test.