Figure 2

Epigenetic regulation of MMP-2 in mesothelioma cells. (a) Reporter gene analysis for the MMP-2 promoter. Relative reporter activity of pGL3-MMP-2 normalized to pGL3-basic is presented as means + s.d. of three independent experiments. (b) Changes in MMP-2 mRNA expression in non-invasive (H28, Meso-4, H2052) and invasive (JMN-1B) mesothelioma cells treated with 5-Aza-2′-deoxycytidine (5Aza-dC) and trichostatin A (TSA). The cells were treated with 5Aza-dC or vehicle for 5 days with drug refreshment every 24 h. The cells were treated with 5Aza-dC and TSA or vehicle for another 24 h. mRNA levels were quantified by qPCR. Relative mRNA levels normalized to GAPDH mRNA are represented as means + s.d. of three independent experiments. (c) Schematic representation of the transcription start site (TSS), CpG dinucleotides, and regions subjected to bisulfite sequencing (BS1 to BS6) in the MMP-2 gene. Methylation of CpG dinucleotides was determined by bisulfite sequencing in non-invasive (H28) and invasive (JMN-1B) cells. (d) Methylation of CpG dinucleotides at BS3 was determined by bisulfite sequencing in normal mesothelial cells (MeT-5A) and mesothelioma cell lines.