Promoter Methylation of RASSF1A Associates to Adult Secondary Glioblastomas and Pediatric Glioblastomas
Figure 2
(a) MGMT and RASSF1A promoter hypermethylation in astrocytomas determined by MSP. PCR products were run in a 2% agarose gel stained with 0.5 μg/mL ethidium bromide. B16: DNA obtained from blood of a normal donor; HN26: pilocytic astrocytoma; HN35: secondary glioblastoma; HN36: primary glioblastoma. m: 1 Kb plus DNA marker; U: unmethylated; M: methylated. C+: DNA methylated with Sss I (CpGenome, Intergen, Edimburgh, UK), positive control for methylation; C−: water. (b) RASSF1A, PTEN, and MGMT promoter hypermethylation in pediatric high-grade astrocytomas determined by MSP. PCR products were run in a 2% agarose gel stained with 0.5 μg/mL ethidium bromide. B27: DNA obtained from blood of a normal donor; T36–T49: DNA from paraffin-embedded pediatric astrocytomas; SW48: colorectal cancer cell line with MGMT promoter hypermethylation, used as a positive control for DNA methylation; C+: Sss I methylated DNA (CpGenome, Intergen, Edimburgh, UK), used as a positive control for methylation; U: unmethylated; M: methylated. (c) PTEN, MGMT, and RASSF1A promoter hypermethylation in astrocytoma cell lines determined by MSP. PCR products were visualized in a 2% agarose gel stained with 0.5 μg/mL ethidium bromide. B23: DNA from blood of a normal donor; GOS3: astrocytoma/oligodendroglioma cell line (grades II/III); U251: glioblastoma multiforme; U118: glioblastoma multiforme; SW1088: anaplastic astrocytoma; SW1783: anaplastic astrocytoma; T98G: glioblastoma multiforme; C+: DNA methylated with Sss I (CpGenome, Intergen, Edimburgh, UK), positive control for methylation; C−: water. Neuroblastoma cell lines Kelly, MC-IXC, Be(2)C, and MHH-NB-11 were included in the study. m: molecular weight DNA marker, 1 Kb Plus DNA ladder (Invitrogen, Life and Technologies, Carlsbad, CA); U: unmethylated; M: methylated. Some other cell lines, different to astrocytoma, appear in the figure.
