Epithelial-Mesenchymal Transition Stimulates Human Cancer Cells to Extend Microtubule-based Invasive Protrusions and Suppresses Cell Growth in Collagen Gel
Figure 4
Growth of three EMT-induced cancer cell lines in three different conditions.
(A) MKN-1 (1.0×104 cells), A549 (0.5×104 cells) and Panc-1 (1.0×104 cells) were incubated in each well containing 1% FBS-containing medium without (Control) or with TGF-ß on 24-well culture plates for 7 days in monolayer culture. After the incubation, the number of cells was measured with a cell counter. Each bar indicates the mean ± SD of the cell numbers in triplicate wells. (B) Cells were cultured at a density of 1×104 cells per well in 24-well suspension culture plates containing 1% FBS-containing medium without (Control) or with TGF-ß for 7 days. The relative number of cells was measured using Dojindo cell counting kit 8. Each bar indicates the mean ± SD of the absorbance values at 485 nm in triplicate wells. (C and D) Cells were inoculated at a density of 7,000 cells per 35-mm dish in 10% FBS-containing soft agar medium without (Control) or with TGF-ß and incubated for 10 (MKN-1) or 14 (A549 and Panc-1) days. After the incubation, cells were stained with p-iodonitrotetrazolium violet, and the number of total colonies (C) and total colony area (D) were determined by Image J and shown as the relative number to the control (100%). Each bar indicates the mean ± SD in triplicate wells. Other experimental conditions are described in Materials and Methods.