Androgen-Regulated Expression of Arginase 1, Arginase 2 and Interleukin-8 in Human Prostate Cancer
Figure 5
Androgens induced Interleukin-8, which in turn promotes ARG1 and ARG2 expression.
Evaluation of the cytokine expression profile of LNCaP cells following R1881 stimulation. A) Conditioned media of LNCaP cells stimulated as previously described were analyzed with a Proteome Profiler™ (R&D Systems). B) Conditioned media of LNCaP cells stimulated over time with either ethanol control (light gray bars) and R1881 (black bars) were analyzed for the production of IL-8 by ELISA. The representative experiment showed was performed with the same conditioned media used for the Proteome Profiler analysis in 5a, (n = 3). C) Quantification of IL-8 secretion by LNCaP cells transfected with siAR and stimulated with R1881 as previously described. Representative experiment is shown, (n = 3). D) Quantification of IL-8 secretion by LNCaP cells transfected with siIL-8 and stimulated with R1881 as previously described. Representative experiment is shown, (n = 3). For 5b and 5c, there was no IL-8 secretion detected in the absence of R1881 stimulation. E) Expression of ARG1 and ARG2 in LNCaP cells following transfection of siIL-8 and R1881 stimulation for 24 hours. Representative experiment is shown, (n = 3). F) LNCaP cells were plated in charcoal-stripped serum supplemented media for 72 hours and for 24 hours in serum-free RPMI. Cells were then stimulated for 72 hours with 10 nM R1881 or with 50, 100 or 250 ng/ml of IL-8 in serum-free RPMI. ARG1 and ARG2 expression levels were detected by Western blot. Representative experiment, (n = 3). Note the induction of both ARG1 and ARG2 at 50 and 100 ng/ml of IL-8 concentration in the absence of R1881.