Abstract
The tumor suppressor genes MGMT and DAPK1 become methylated in several cancers including diffuse large B-cell lymphoma (DLBCL). However, allelic methylation patterns have not been investigated in DLBCL. We developed a fast and cost-efficient method for the analysis of allelic methylation based on pyrosequencing of methylation specific PCR (MSP) products including a SNP. Allelic methylation patterns were reliably analyzed in standards of known allelic methylation status even when diluted in unmethylated DNA to below 1% methylation. When studying 148 DLBCL patients MGMT and DAPK1 methylation was observed in 19% and 89%, respectively, and among methylated and heterozygous patients 29% and 55%, respectively, were biallelically methylated. An association between the T-allele of the rs16906252 SNP and MGMT methylation was observed (p-value=0.04), and DAPK1 methylation of the A-allele was associated with shorter overall survival (p-value=0.006). In future cancer research allelic MSP-pyrosequencing may be used to study a wide range of other loci.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adult
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Aged
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Aged, 80 and over
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Alleles
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Antibodies, Monoclonal, Murine-Derived
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Antineoplastic Combined Chemotherapy Protocols
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Cell Line
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Cyclophosphamide
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DNA Methylation*
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DNA Modification Methylases / genetics*
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DNA Repair Enzymes / genetics*
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Death-Associated Protein Kinases / genetics*
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Doxorubicin
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Female
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Genotype
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Humans
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Lymphoma, Large B-Cell, Diffuse / drug therapy
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Lymphoma, Large B-Cell, Diffuse / genetics*
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Lymphoma, Large B-Cell, Diffuse / mortality
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Lymphoma, Large B-Cell, Diffuse / pathology
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Male
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Middle Aged
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Neoplasm Staging
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Polymorphism, Single Nucleotide
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Prednisone
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Rituximab
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Sensitivity and Specificity
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Sequence Analysis, DNA
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Treatment Outcome
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Tumor Suppressor Proteins / genetics*
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Vincristine
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Young Adult
Substances
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Antibodies, Monoclonal, Murine-Derived
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R-CHOP protocol
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Tumor Suppressor Proteins
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Rituximab
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Vincristine
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Doxorubicin
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Cyclophosphamide
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DNA Modification Methylases
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MGMT protein, human
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DAPK1 protein, human
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Death-Associated Protein Kinases
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DNA Repair Enzymes
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Prednisone