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Fig. 5 | Molecular Cancer

Fig. 5

From: 3-BrPA eliminates human bladder cancer cells with highly oncogenic signatures via engagement of specific death programs and perturbation of multiple signaling and metabolic determinants

Fig. 5

Aberrant activities of Akt and p44/42 MAPK signaling pathways critically modulate bladder cancer cell sensitivity to 3-BrPA. (a-b) Representative (three independent experiments) Western blotting profiles of whole-cell protein extracts derived from T24 cells, grown at ~60 % confluency and treated with the indicated doses of 3-BrPA for 24 h, in the absence or presence (pre-incubation for 2 h) of 100 μM LY294002 (a) or 100 μM U0126 (b). The proteins examined were p-Akt-Ser473, Akt (a), p-p44/42 MAPK-Thr202/Tyr204 and p44/42 MAPK (b), while Actin was used as molecule of reference. Brackets and asterisks denote the high MW forms of Akt (a) and p44/42 MAPK (b) kinases. (c-e) MTT cytotoxicity assays of T24 cells, seeded at low confluency and exposed to the indicated doses of 3-BrPA for 24 h, in the absence or presence (pre-incubation for 2 h) of 100 μM LY294002 (c), 100 μM U0126 (d) and 1 mM AICAR (e). LY294002, U0126 and AICAR remained in the growth medium with half of their initial respective concentrations for 24 h more, post-pre-incubation. Stock solutions for all three reagents (LY294002, U0126 and AICAR) were prepared in DMSO. MTT viability rates obtained from the cocktail of 3-BrPA with each kinase inhibitor (LY294002 and U0126), or activator (AICAR), were compared to the respective ones of 3-BrPA alone and, after their normalization with values derived from inhibitor/activator only, versus its cognate solvent, they were presented in fold (x) of inhibitor/activator-induced cell survival increase. (c-e) Data are reported as mean ± standard deviation of triplicates of three independent experiments. *P < 0.001. (f-g) MTT cytotoxicity assays of RT112 (f) and TCCSUP (g) cells, grown at ~60 % confluency and treated with the indicated doses of 3-BrPA for 24 h (also, see Additional file 10: Table S2). (f-g) Results are denoted as mean ± standard deviation of triplicates of, at least, three independent experiments. *P < 0.001

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