Alcohol increases p53 transcriptional activity and induces cell cycle arrest.

<p><b>A)</b> Relative mRNA levels of <i>TP53</i> are shown from MCF-7 cells treated with alcohol (0, 0.2%, or 0.4% v/v) for 6 hours. Values are presented as the mean ± S.E. <b>B)</b> MCF-7 cells transiently transfected with the p53 luciferase reporter plasmid (MCF-7/p53-luc cells) were treated with alcohol (0, 0.2%, or 0.4% v/v) for 2 hours. Then, the cell lysates were prepared for reporter assays. The luciferase activity relative to the control for each sample is shown. Values are presented as the mean ± S.E. (**p<0.01). <b>C)</b> MCF-7 cells were treated with various doses of alcohol (0, 0.1%, 0.2%, or 0.4% v/v) for 6 hours. mRNA levels of p21 and Bax were measured using qPCR. Relative fold changes for the alcohol-treated samples are displayed as compared to the respective control. Values are presented as the mean ± S.E. (**p<0.01). <b>D)</b> MCF-7 cells were treated with alcohol (0, 0.1%, 0.2%, 0.4%, or 0.8% v/v) for 6 hours and then harvested for protein expression. p21 and Bax protein levels were determined by Western blot analysis. <b>E)</b> MCF-7 cells were exposed to alcohol (0, 0.2%, 0.4%, or 0.8% v/v) for 24 hours before cells were fixed and prepared for FACS analysis as described in the <i>Materials and Methods</i>. <b>F)</b> The graph depicts the average percentage of cells (± S.E.) in G0/G1, S, and G2/M phase (*p<0.05; **p<0.01 as compared to the control) from three replicate experiments.