Nrf2 Expression Is Regulated by Epigenetic Mechanisms in Prostate Cancer of TRAMP Mice
Figure 4
The expression of Nrf2 and NQO-1 in TRAMP cell lines was correlated with CpG islands methylation status and could be restored by 5-aza/TSA treatment.
TRAMP C1 and C3 cells were treated with 2 µM 5-aza, 200 nM TSA, or 1 µM 5-aza plus 100 nM TSA for 60 hrs or 48 hrs followed by incubation in the presence of 5 µM tBHQ for further 12 hrs. After treatments, the cells were harvested for total protein or RNA extractions. (A) the mRNA levels of Nrf2 and NQO1 were determined by RT-PCR, with GAPDH serving as internal control, and the results from 3 independent experiments were quantified by densitometry and the results are shown in B and C. (D) the protein levels of Nrf2, NQO1, MBD2, H3K9me3 and H3Ac were determined by Western blotting, with actin as a loading control. Each experiment was repeated at least twice with similar results.