FIGURE 3.

Cytostatic effects after combined treatment with metformin, diclofenac and diflunisal on AML cells. 3 × 10⁴cells/0.2 mL medium (A) U937 cells or (B) THP-1 cells or (C) primary blasts were seeded in 96-well plates with 1 mM metformin, 0.1 mM diclofenac and 0.1 mM diflunisal alone or in the indicated combinations. ³H-thymidine incorporation was determined after 24 h. In a second set of experiments we increased the diflunisal concentration. (D) THP-1 cells or (E) primary blasts were incubated with 0.1 mM up to 0.8 mM diflunisal plus low dose 0.1 mM diclofenac for 24 h. (F) alternatively, the incubation period of primary blasts was prolonged to 48 h. (G–I) in another set of experiments the triple combination with diclofenac and 1 mM metformin was used. Proliferation under single drug treatment or the various drug combinations was normalized to respective controls and is shown for (J) THP-1 cells and primary blasts treated for (K) 24 h or (L) 48 h. data are shown as mean ± SEM (n ≥ 3), ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001 significance was calculated by ANOVA and post hoc Holm-Sidak’s multiple comparisons test.