Figure 1.
Reduction of DNMT1 promotes EMT induction in PCa cells.
PCa cells (PC3 or DU145) (1 × 105) were seeded in 6-well culture plates. The cells were treated by vehicle or 5-Aza (0-20 μM) for 4 days.
(A) mRNA levels of DNMT1 expression in vehicle- or 5-Aza (5 μM)–treated PCa cells (PC3 or DU145) as quantified by real-time PCR. The human prostate epithelial cell RWPE-1 was used as a control. (B) Protein levels of DNMT1 following vehicle or 5-Aza (5 μM) treatment in PCa cells (PC3 or DU145) as quantified by Western blots. (C) mRNA levels of EMT markers in vehicle- or 5-Aza (5 μM)–treated PCa cells (PC3 or DU145) as quantified by real-time PCR. (D) E-Cadherin or N-Cadherin expression in vehicle- or 5-Aza (5 μM)–treated PCa cells (PC3 or DU145) as detected by immunofluorescence staining. Blue, DAPI nuclear stain. Bar = 20 μm. (E) EMT markers in vehicle- or 5-Aza (5 μM)–treated PCa cells (PC3 or DU145) as quantified by Western blots. (F) Migration of vehicle- or 5-Aza (5 μM)–treated PCa cells toward the chemoattractant CXCL12 was performed in Transwell plates. Percent migration was set at 100% for the initial cell numbers in top chamber. The data in A, C, and F are representative of mean ± SD (n = 6). P values were calculated by Student's t test.