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Figure 5 | Journal of Experimental & Clinical Cancer Research

Figure 5

From: p38α MAPK-mediated induction and interaction of FOXO3a and p53 contribute to the inhibited-growth and induced-apoptosis of human lung adenocarcinoma cells by berberine

Figure 5

BBR-induced inhibition of cell growth and induction of apoptosis were dependent on p53 and FOXO3a protein expression in A549 cells. A-B, A549 cells were treated with Pifithrin-α (10 μM) for 2 h before exposure the cells to BBR (25 μM) for an additional 24 h followed by measuring the p53 protein expression (A). GAPDH was used as internal control (A). And cell cycle was analyzed by flow cytometry after propidium iodide (PI) staining (B). The bar graphs represent the mean ± SD of p53/GAPDH or relative percentage of cell cycle phases of three independent experiments. C-D, Cells were transfected with control or p53 or FOXO3a siRNAs with lipofectamine 2000 reagent for 24 h, followed by exposure the cells to BBR (25 μM) for an additional 24 h. Afterwards, the cell proliferation was detected using MTT assays. The expression of p53 and FOXO3a protein was determined by Western blot. The bar graphs represent the mean ± SD of p53/GAPDH and FOXO3a/GAPDH of three independent experiments. E, Cells were transfected with control or FOXO3a siRNAs (50 nM each) for 24 h before exposing the cell to BBR for an additional 24 h. Afterwards, cell apoptosis was determined by flow cytometry assays. F, Cells were transfected with control (pEGFP-N1) or FOXO3a expression vector (FOXO3a-pEGFP) for 24 h before exposing the cells to BBR for an additional 24 h. Afterwards, the expression of FOXO3a protein and apoptosis were detected by Western blot and flow cytometry, respectively. Data are expressed as a percentage of total cells. Values in bar graphs were given as the mean ± SD from three independent experiments performed in triplicate. *indicates significant difference as compared to the untreated control group (P < 0.05). **Indicates significant difference from BBR treated alone (P < 0.05).

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