Figure 5

Inhibition of cell growth and migration of P6C cells by compound 1. (a) Compound 1 inhibited the cell growth of P6C cells. P6C cells were treated with compound 1 for 24 hours with increasing doses as indicated. The MTT viability assay was used to measure the cytotoxic effects of compound 1. (b,c) Compound 1 inhibited the colony forming ability of P6C cells. Approximately 150 P6C cells were seeded in the wells of a 6-well plate, and compound 1 was added at different concentrations. After two weeks, the cells were fixed and stained with Coomassie Brilliant Blue R250 and counted. Representative images of the plates are shown (b), and the number of colonies is shown in the bar graph (c). (d) Compound 1 inhibited the sphere-forming ability of P6C cells. Approximately 200 P6C cells were seeded in 0.3% soft agar for the anchorage-independent sphere-forming assay. After three weeks, the spheres were fixed, stained and counted. The number of spheres is shown in the bar graph. (e) Compound 1 inhibited the migration ability of P6C cells. P6C cells were seeded in the upper layer of the well. Migrated cells on the bottom side of transwell cultures were fixed and stained. Images were taken in 5 random microscopic fields using a light microscope. In addition, the migrated cells were quantified with ImageJ. All the experiments were performed in triplicate and repeated three times with similar results. Bars represent the mean ± SD.