Figure 4
From: Synthesis and mechanisms of action of novel harmine derivatives as potential antitumor agents
Compound 3c induced cell apoptosis through suppression of PI3K/AKT signaling pathway in cancer cells.
(A) Luciferase activity of NRF2, HSF1, stat3, PI3K and NFκB signaling pathway reporter. HCT116 cells were transfected with the indicated pathway reporter for 24 h, treated with 4 μM of compound 3c for another 24 h and then subjected to dual-luciferase assay. (B) Dose-dependent reduction of PI3K reporter activity. The cells were incubated with the indicated concentration of compound 3c for 24 hours. (C) Dose- and time- dependent inhibition of the levels of phosphorylated AKT and ERK. HCT116 cells were treated with the indicated concentration of compound 3c for 48 hours or with 8 μM compound 3c for 8, 16, 32 and 48 hours. (D) Luciferase analysis of PI3K/AKT reporter activities and western blotting for measurment of pAKT levels in HepG2, Hela and A549. For luciferase assay, cells were treated with 8 μM compound 3c for 18 hours. (n = 3, *P < 0.05 versus DMSO). For western blotting, cells were treated with the indicated concentration of compound 3c for 72 h. (E) LY294002 inhibited the phosphorylation level of AKT. HCT116 cells were treated with 8 μM compound 3c alone for 24 hours or in combination with pretreatment with 50 μM LY294002 for 4 hours. (F) The protein levels of PARP, caspase-3 and cleaved caspase-3 in HCT116 cells treated with 8 μM compound 3c alone or in combination with 50 μM LY294002 for 4 h. The data represents the means ± SDs. GAPDH was used as an internal control.
