Figure 2.

SDC-1 up-regulation by n-3 PUFA in prostates of Pten^P-/- mice/cells and in human prostate cancer cells. (A) Protein extracts from the prostates of 8-week-old Pten^P+/+ and Pten^P-/- mice fed n-3 PUFA or n-6 PUFA diets were dialyzed and treated with heparinase III plus chondroitin ABC lyase for 18 hours at 37°C. SDC-1 was measured by Western blot assay and normalized to β-actin. Values representing the mean and SD (n = 3) for SDC-1/β-actin ratios with different letters are significantly different (P < .05). (B) Cultured prostate cells from Pten^P+/+ and Pten^P-/- mice were harvested at approximately 70% confluence, and (C) Pten^P-/- prostate cells were incubated with n-3 or n-6 PUFA-enriched LDL (100 µg/ml) for 8 hours. Total RNA was isolated, and SDC-1 mRNA was determined by real-time PCR. Values expressed relative to controls (n = 3) with different letters are significantly different (P < .05). (D) Human prostate cells (normal prostate epithelial cells [PrEC] and prostate cancer cells [PC3, LNCaP, and DU145]) were harvested at approximately 70% confluence. Total RNA was isolated, and SDC-1 mRNA was determined by real-time PCR. Values, expressed relative to controls (n = 3), with different letters are significantly different (P < .05).