Fig. 3. Effect of GA or CDDP on survival and progression of A549 cells. (A) A549 cells were seeded at a density of 500 cells per well in 6-well plates, and treated with NC (0.1% DMSO), CDDP (12 μM) and GA (75 μM). Colony formation assay was performed to measure the colony forming ability. The number of colonies was quantified using the Image J program. (B) A549 cells were seeded at a density of 500 cells/25 μL/droplet on the lid of petri dish, in appropriate media containing the test compounds, and incubated for 6 days. The size of tumor spheroids generated was determined using the Image J program. A549 cells were seeded at a density of 6×105 cells in cell culture dishes and treated with NC (0.1% DMSO), CDDP (12 μM) and GA (75 μM) for 48 h. (C) The expressions of cell cycle related proteins such as Cyclin D1, Cyclin E1, p21 and p27 were confirmed by Western blot assay. (D) The expression levels of each protein were normalized to GAPDH and quantified. The representative images were obtained from at least three repeated experiments. Data are expressed as means ± SD from three independent experiments. *p<0.05 vs. NC (0.1% DMSO).
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