Figure 6From: Estrogen receptor beta as a novel target of androgen receptor action in breast cancer cell linesER beta silencing counteracts the effects of mibolerone on p21, cyclin D1 expression and cell proliferation. (A) RNA was extracted from MCF-7 cells treated with vehicle (−) or Mib 10 nM for 48 hours in the presence of a non-specific or an ER beta siRNA, reverse transcribed and cDNA was subjected to Real-Time RT-PCR for p21 and cyclin D1 mRNA expression. Each sample was normalized to GAPDH mRNA content. Data represent the mean ± S.D. of values from three separate RNA samples expressed as the percentage of the control (−) assumed to be 100%. (B) Western blot analysis for p21 and cyclin D1 expression in cells treated with vehicle (−) or Mib for 48 hours in the presence of a non-specific or an ER beta siRNA. GAPDH was used as the loading control. The histograms represent the mean ± S.D. of three independent experiments in which band intensities were evaluated in terms of optical density arbitrary units and expressed as the percentage of the control assumed to be 100%. (C, D) MTT assays in MCF-7 and ZR-75 cells transfected and treated as indicated. Results are expressed as fold change ± S.D and are representative of three different experiments each performed in triplicate. *, P <0.01 compared to vehicle-treated cells. **, P <0.01 ER beta siRNA-transfected cells compared to Mib-treated cells. ER, estrogen receptor; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.Back to article page