5-Aza/E2 inhibits TAM-Wd cell proliferation.

A. TAM-Wd and MCF-7 cell concentration response to E2 challenge (1×10⁻¹² M to 1×10⁻⁷ M) ±5-Aza (1×10⁻⁶ M). Cell counts were taken on day 7 of culture. Data shown represent E2-treated cell counts as a percentage of non-E2 treated control cells. Cell counts are significantly lower in 5-Aza treated TAM-Wd cells, co-treated with E2 at a concentration of 1×10⁻¹⁰ M and greater (*p<0.017). B. TAM-Wd cell concentration response to E2 challenge (1×10⁻¹² M to 1×10⁻⁷ M) +5-Aza (1×10⁻⁶ M) ±TAM (1×10⁻⁷ M). Cell counts were taken on day 7. Data shown represent E2-treated cell counts as a percentage of non-E2 treated control cells. The co-addition of TAM to 5-Aza treated TAM-Wd cells significantly changes the effect of E2 from a concentration of 1×10⁻⁹ M and greater (*p<0.026). C. Anchorage-dependent proliferation assay of TAM-Wd cells treated with E2 (1×10⁻⁹ M) ± TAM (1×10⁻⁷ M) in the presence of 5-Aza (1×10⁻⁶ M) for 14 days. The data shown represent actual cell number/well recorded over 3 independent experiments. By day 14, there are significantly more cells in the 5-Aza and 5-Aza/E2/TAM treated cells compared to TAM-Wd +5-Aza/E2 treated cells (*p<0.001). D. Cell cycle analysis using flow cytometric analysis of propidium iodide-stained TAM-Wd cells. Data represent percentage of cell in each phase relative to the total population.