Fig. 2

GR liganding inhibits ER-mediated proliferative gene expression. a MCF-7 cells were treated with vehicle (ETOH), 100 nM Dex, 10 nM E2, or Dex/E2 for 4 h and global gene expression was evaluated by microarray followed by Ingenuity Pathway Analysis (IPA). Proliferation and cell cycle pathways were identified as those with the lowest p values; pathway activation z-score is also shown. Dex/E2 treatment consistently demonstrated relatively decreased gene expression pathway activation (z-score) compared to E2-alone. b Genes common to cell proliferation pathways in 2A (CCND1, CDK2, and CDK6) were assessed by qRT-PCR in independent experiments. All genes demonstrated reduced gene expression by 24 h following Dex treatment; with the exception for CDK6 in MCF-7 cells, all genes demonstrated significant reduction in gene expression following SGRM treatment (*p < 0.05, **p < 0.01, ****p < 0.0001, two-way ANOVA, Tukey’s post hoc test, n = 3, ±SD). C) MCF-7 and T-47D wild-type ER+ cells were treated with vehicle (Veh, ETOH), 10 nM E2, 1 μM C134/E2 (134/E2), or 1 μM C335/E2 (335/E2) for 3 days. Cyclin D1, ERα, and β-actin were immunoblotted. E2 treatment led to an increase in Cyclin D1 protein expression, while C335 led to relatively decreased Cyclin D1 protein expression in MCF-7 cells. C134 led to a more pronounced decrease in Cyclin D1 protein in T-47D cells. Densitometry analysis is shown below the Cyclin D1 bands (relative to Veh). ERα protein expression was unchanged following combined E2/SGRM treatment when compared to either E2-alone or vehicle (Veh)