Figure 3

Hormone activates the VEGF promoter in two different cell lines, LNCaP and 22Rv1. (A) Schematic diagram of the VEGF promoter showing locations of predicted ARE binding sites (grey boxes) in relation to 5’ termini of luciferase reporter constructs V411 and V2274 [16]. (B) 22Rv1 cells were transfected with the 411 bp VEGF promoter construct (V411) in the presence or absence of different concentrations of R1881 (0, 0.05nM, 0.5nM, and 5nM) for 48hrs. Luciferase assays of cell lysates were performed and fold activation was determined as described in text. (C) 22Rv1 cells were transfected with V411 and treated with 0nM R1881, or 5nM R1881 with 0μM casodex, or 5nM R1881 with 10μM casodex for 48 hrs. (D) LNCaP cells were transfected and treated as per (C). (E) 22Rv1 cells were transfected with the 2274 bp VEGF promoter construct (V2274) and treated as per (C). Luciferase activity is shown relative to average normalized luciferase activity in the absence of hormone. Experiments were repeated three times in triplicate. Significance was determined by Student’s t-test (*p<0.05, ** p<0.01, ***p<0.001).