MicroRNA-596 acts as a tumor suppressor in gastric cancer and is upregulated by promotor demethylation

Figure 5 MicroRNA-596 is regulated by epigenetic mechanisms in gastric cancer. A: Methylation-specific PCR (MSP) analysis of microRNA-596 (miR-596) promoter region methylation in gastric cancer (GC) cell lines. B: Quantitative real-time PCR (qRT-PCR) and MSP analysis of miR-596 expression after treatment with 5-Aza-2’-deoxycytidine (5-Aza-dC) (2.5, 5, or 10 µmol/L) in MKN-45 and MGC-803 cells, ^aP < 0.05, ^bP < 0.01 vs control cells. C: qRT-PCR and Western blot analysis of PRDX1 expression after treatment with 5-Aza-dC (2.5, 5 or 10 µmol/L) in MKN-45 and MGC-803 cells, ^aP < 0.05, ^bP < 0.01 vs control cells. D: The promoter sequence of miR-596 after treatment with sodium bisulfate. Unmethylation of cytosine was transformed to uracil, but methylated cytosine remained unchanged. 1: DNA sequence of miR-596 promoter; 2: Sequence after sodium bisulfate modification; U: Unmethylation lane; M: Methylation lane; PRDX1: Peroxiredoxin 1; MiR-596: MicroRNA-596; 5-Aza-dC: 5-Aza-2’-deoxycytidine.