Figure 3.

Exposure to alcohol resulted in the activation of the MAPK pathway and increased cell proliferation. (A) T47D cells were incubated in the presence or absence of 10 μM U0126, a MEK1/2 inhibitor, for 0.5, 1, 3, 6, 12 and 24 h. Equal amounts of proteins were obtained from the cells and were analyzed using western blotting to detect p-MEK1/2 (S217/S2221), p-ERK1/2 (T202/Y204) and p-MSK1 (T581). Phosphorylated MEK was decreased at 0.5, 1 and 3 h in the T47D cells exposed to alcohol in the presence of U0126. Phosphorylated ERK1/2 was decreased at 0.5, 1, 3, 6, 12 and 24 h in the T47D cells treated with alcohol in the presence of U0126. Phosphorylated MSK was decreased at 12 and 24 h in the T47D cells treated with alcohol in the presence of U0126. (B) The T47D cells were exposed to 100 mM alcohol and/or 10 μM U0126 for 12 and 24 h; the cell proliferation was evaluated, and the proliferation of the cells was measured using WST-1. The T47D cells treated with alcohol demonstrated an increase in cell proliferation and the U0126-treated cells showed a decrease compared with the untreated cells. However, exposure to both alcohol and U0126 induced an increase in cell proliferation when compared to the U0126-treated cells. The values are presented as means ± SEM (n=3). ^*Significantly different from the NC group. A one-way ANOVA followed by Tukey’s HSD post-hoc test was used to evaluate the statistical significance of the differences. NC, untreated control.