T877A mutant AR mediates the stimulatory effects of genistein.

<p>A: Graphic presentation of the effect of combined treatment with R1881 (1nmol/L) and Casodex (100nmol/L) or genistein (1µmol/L) and Casodex (100nmol/L) on LNCaP cell growth measured by MTS assay. Results in the graph represent the mean ± SD of three experiments each in triplicate. *p < 0.05, **p <0.01, for comparisons with the groups treated with R1881 or genistein, only. B: Evaluation of genistein–AR binding <i>in </i><i>silico</i>. Representative figure showing conformational changes of the WT-AR and the T877A mutant AR after molecular docking with genistein <i>in </i><i>silico</i> using the GOLD v5.0.1 modeling program. Different domains of the AR are presented in various colors. Inset, genistein bound to the ligand-binding domain (LBD) of AR-receptor. Arrows point to different conformational changes and loop movements in WT-AR and T877A mutant AR in response to genistein docking. C: Geometric optimization, and tautomeric forms of genistein created at pH 7.5 ± 0.5 using EPIK. The yellow is for the WT-AR, and the green is for the T877A mutant AR. D and E: Evaluation of genistein–AR binding using ³[R1881] competitive binding assay. Histogram shows the binding of genistein with AR in LAPC4 (WT-AR) and LNCaP (mutant AR) cells as assessed by radiolabeled cell-based competitive binding assay as described in Materials and Methods. Results in the graph represent the mean ± SD of three experiments. *p < 0.05, **p <0.01, compared with control.