Mitsuhiro Anan, Rolando Del Maestro, Nobihiro Hata, Minoru Fujiki
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10125-GGE-10 EVALUATION OF MGMT PROMOTER METHYLATION STATUS OF GBM CELL LINE CLONAL POPULATION.
Abstract In this study, the epigenetic heterogeneity of GBM was investigated by evaluating the methylation status of the O6-methylguanine methyltransferase (MGMT) promoter in individual clones of a single cell derived from GBM cell lines. Cell lines, U251 and U373, were used for the experiments from the Brain Tumour Research Centre of the Montreal Neurological Institute. To evaluate the methylation status of the MGMT promoter, pyrosequencing and methylation-specific PCR (MSP) were used. A total of 12 U251 and 12 U373 clones were isolated. The methylation status of 83 of 97 CpG sites in the MGMT promoter were evaluated by pyrosequencing, and 11 methylated CpG sites and 13 unmethylated CpG sites were evaluated by MSP. The results demonstrate tumor heterogeneity among individual clones derived from a single GBM cell.
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