Alisol B acetate induces apoptosis of SGC7901 cells via mitochondrial and phosphatidylinositol 3-kinases/Akt signaling pathways

Figure 1 Effect of alisol B acetate on survival of SGC7901 cells. Cells were incubated in the absence or presence of various concentrations of alisol B acetate for 24 h (A), or at 30 &mgr;mol/L for different incubation times (B). ^aP < 0.05, ^bP < 0.01 vs control group (unpaired Student’s t test).

Figure 2 Morphology of SGC7901 cells exposed to alisol B acetate for different concentrations observed under phase-contrast microscope. A: Controls; B-D: SGC7901 cells were treated with 30, 50 and 70 &mgr;mol/L alisol B acetate for 24 h.

Figure 3 Morphological changes observed under electronic microscope. A: Control; B-D: Cells were incubated with 30 &mgr;mol/L alisol B acetate for 24, 48 and 72 h.

Figure 4 Effect of alisol B acetate on cell cycle distribution of SGC7901 cells. SGC7901 cells were treated with 0.1% DMSO (A) or with 30 &mgr;mol/L alisol B acetate for 24 h (B), 48 h (C), or 72 h (D). The cells were collected sequentially and stained with PI and analyzed by FCM.

Figure 5 FCM analysis of mitochondrial membrane potential in human SGC7901 cells with 30 &mgr;mol/L alisol B acetate for various time periods. The zero concentration was defined as control. The percentage of cells stained with Rh-123 was determined by FCM as described in the Materials and Methods. A: Control; B-E: 12, 24, 48 and 72 h.

Figure 6 Effect of Alisol B acetate on the expression of Bcl-2, Bax, Apaf-1, cleaved caspase-3 and caspase-9. SGC7901 cells were treated with 30 &mgr;mol/L alisol B acetate for 12, 24, 48 and 72 h. The cells were collected and lysed. Western blotting analysis was conducted and probed with antibodies to Bcl-2, Bax, Apaf-1, caspase-3 and caspase-9. Lanes (from left to right): Control cells; 12 h; 24 h; 48 h; 72 h.

Figure 7 Effect of alisol B acetate on PI3K/Akt activation in SGC7901 cells. SGC7901 cells were treated with 0.1% DMSO (A) or 30 &mgr;mol/L alisol B acetate for 12, 24, 48 and 72 h. Western blotting analysis for PI3K, t-Akt and P-Akt was performed using specific antibodies with β-actin as a loading control.