Figure 1.

Da0324 inhibits proliferation and induces apoptosis and autophagy in gastric cancer cells. A. Gastric cancer cell lines (BGC823, SGC7901 and KATO III) were treated with the indicated concentration of Da0324 for 24 h, 48 h, or 72 h. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) and Growth inhibition rates of Da0324 on GC cells were in comparison with untreated cells. B. BGC823, SGC7901 and KATO III cells were treated with 0, 0.5, 1, 2, 4, 6, 8, 10, or 12 mM Da0324 for 48 h, and cell viability was measured by CCK-8 assays. The half maximal inhibitory concentration (IC50) values were calculated with GraphPad statistical software. C. Clonogenic assay showed the effects of Da03224 treatment on clonogenic formation in gastric cancer cells. BGC823, SGC7901 and KATO III cells were treated with Da0324 (1 or 2 µM) for 48 h, then cultured for two weeks in complete medium and the colony density calculated. Representative images of clonogenic assay (left panel) and quantitative analysis (right panel). All data are representative of three independent experiments and are presented as the means ± SD. *, P < 0.05; **, P < 0.01. D. BGC823, SGC7901 and KATO III cells were treated with 4 μM Da0324 for 48 h. Cells were fixed and stained with Annexin V/PI and apoptosis was analyzed by flow cytometry. Representative dot plots of Annexin V/PI staining are shown in the left panel, and quantitative data are presented in the right panel. All data are representative of three independent experiments and are presented as the means ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001. E. BGC823 and SGC7901 cells were treated with 4 μM Da0324 or control for 48 h and then harvested for analysis by western blot using P53, Bcl-2, p-mTOR, mTOR, P62, and LC3B antibodies. GAPDH or β-actin was used as an internal control.