Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing

Lasse Sommer Kristensen; Marianne Bach Treppendahl; Fazila Asmar; Mia Seremet Girkov; Helene Myrtue Nielsen; Tina Ellegaard Kjeldsen; Elisabeth Ralfkiaer; Lise Lotte Hansen; Kirsten Grønbæk

doi:10.1038/srep02789

Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing

Lasse Sommer Kristensen, Marianne Bach Treppendahl, Fazila Asmar, Mia Seremet Girkov, Helene Myrtue Nielsen, Tina Ellegaard Kjeldsen, Elisabeth Ralfkiaer, Lise Lotte Hansen, Kirsten Grønbæk

Department of Biomedicine - Research and Education, Bartholin Building

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

30 Citations (Scopus)

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.

Original language	English
Journal	Scientific Reports
Volume	3
Pages (from-to)	2789
ISSN	2045-2322
DOIs	https://doi.org/10.1038/srep02789
Publication status	Published - 2013

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@article{5db296393c804fee83d062165499fb8d,

title = "Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing",

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.",

author = "Kristensen, {Lasse Sommer} and Treppendahl, {Marianne Bach} and Fazila Asmar and Girkov, {Mia Seremet} and Nielsen, {Helene Myrtue} and Kjeldsen, {Tina Ellegaard} and Elisabeth Ralfkiaer and Hansen, {Lise Lotte} and Kirsten Gr{\o}nb{\ae}k",

year = "2013",

doi = "10.1038/srep02789",

language = "English",

volume = "3",

pages = "2789",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

}

Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing. / Kristensen, Lasse Sommer; Treppendahl, Marianne Bach; Asmar, Fazila et al.
In: Scientific Reports, Vol. 3, 2013, p. 2789.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing

AU - Kristensen, Lasse Sommer

AU - Treppendahl, Marianne Bach

AU - Asmar, Fazila

AU - Girkov, Mia Seremet

AU - Nielsen, Helene Myrtue

AU - Kjeldsen, Tina Ellegaard

AU - Ralfkiaer, Elisabeth

AU - Hansen, Lise Lotte

AU - Grønbæk, Kirsten

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

U2 - 10.1038/srep02789

DO - 10.1038/srep02789

M3 - Journal article

C2 - 24071855

SN - 2045-2322

VL - 3

SP - 2789

JO - Scientific Reports

JF - Scientific Reports

ER -

Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing

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