Control of DNA methylation at the CpG island in the ST3Gal II promoter in prostate cancer cells.
(A) The CpG island in the ST3Gal II p1 promoter and the location of the MSP primers. The vertical bars represent CpG sites and TSS represents the transcriptional start site. (B–D) The MSP analyses of the CpG island of ST3Gal II. DNA was isolated from LNCap cells treated with 5-azadC (0–50 µM) for 120 h (B), castration-resistant prostate cancer cell lines (PC3 and DU145) or LNCap cells treated with or without 100 nM testosterone for 120 h (C) or LNCap cells treated with or without 100 nM testosterone simultaneously with or without 10 µM bicalutamide for 120 h (D). Then, the DNA was treated with sodium bisulfite, and finally amplified with primers specific for the unmethylated (USP) or the methylated (MSP) form of the CpG island in the ST3Gal II promoter (M, methylated control; UA, unmethylated control A; UB, unmethylated control B). The MSP analyses were repeated 3 times with the same results, and a representative image is shown in the figures.