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Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence

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Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence. / Krais, Annette M; Kliem, Christian; Arlt, Volker M; Schmeiser, Heinz H.

In: Electrophoresis, Vol. 40, No. 11, 01.06.2019, p. 1535-1539.

Research output: Contribution to journalArticle

Harvard

Krais, AM, Kliem, C, Arlt, VM & Schmeiser, HH 2019, 'Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence', Electrophoresis, vol. 40, no. 11, pp. 1535-1539. https://doi.org/10.1002/elps.201800495

APA

Krais, A. M., Kliem, C., Arlt, V. M., & Schmeiser, H. H. (2019). Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence. Electrophoresis, 40(11), 1535-1539. https://doi.org/10.1002/elps.201800495

Vancouver

Krais AM, Kliem C, Arlt VM, Schmeiser HH. Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence. Electrophoresis. 2019 Jun 1;40(11):1535-1539. https://doi.org/10.1002/elps.201800495

Author

Krais, Annette M ; Kliem, Christian ; Arlt, Volker M ; Schmeiser, Heinz H. / Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence. In: Electrophoresis. 2019 ; Vol. 40, No. 11. pp. 1535-1539.

Bibtex Download

@article{4feaab3e18524971a62c2847f69a663e,
title = "Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence",
abstract = "Methylating substances alter DNA by forming N3-methylthymidine (N3mT), a mutagenic base modification. To develop a sensitive analytical method for the detection of N3mT in DNA based on capillary electrophoresis with laser-induced fluorescence detection (CE-LIF), we synthesized the N3mT-3'-phosphate as a chemical standard. The limit of detection was 1.9 amol of N3mT, which corresponds to one molecule of N3mT per 1,000 normal nucleotides or 0.1%. With this method, we demonstrated that the carcinogenic nitrosamine N'-nitrosonornicotine (NNN) induced N3mT in the human lung cancer cell line A549. Treatment with NNN also caused an elevated degree of 5-hydroxymethylcytidine (5hmdC) in DNA, while the methylation degree (i.e. 5-methylcytidine; 5mdC) stayed constant. According to our data, NNN could, via yet unknown mechanisms, play a role in the formation of N3mT as well as 5hmdC. In this study we have developed a new sensitive analytical method using CE-LIF for the simultaneous detection of the three DNA modifications, 5mdC, 5hmdC and N3mT. This article is protected by copyright. All rights reserved.",
keywords = "5-hydroxymethylcytosine, Capillary electrophoresis with laser-induced fluorescence, DNA methylation, N3-methylthymidine, Nitrosamines",
author = "Krais, {Annette M} and Christian Kliem and Arlt, {Volker M} and Schmeiser, {Heinz H}",
note = "{\textcopyright} 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.",
year = "2019",
month = jun,
day = "1",
doi = "10.1002/elps.201800495",
language = "English",
volume = "40",
pages = "1535--1539",
journal = "Electrophoresis",
issn = "0173-0835",
number = "11",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence

AU - Krais, Annette M

AU - Kliem, Christian

AU - Arlt, Volker M

AU - Schmeiser, Heinz H

N1 - © 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

PY - 2019/6/1

Y1 - 2019/6/1

N2 - Methylating substances alter DNA by forming N3-methylthymidine (N3mT), a mutagenic base modification. To develop a sensitive analytical method for the detection of N3mT in DNA based on capillary electrophoresis with laser-induced fluorescence detection (CE-LIF), we synthesized the N3mT-3'-phosphate as a chemical standard. The limit of detection was 1.9 amol of N3mT, which corresponds to one molecule of N3mT per 1,000 normal nucleotides or 0.1%. With this method, we demonstrated that the carcinogenic nitrosamine N'-nitrosonornicotine (NNN) induced N3mT in the human lung cancer cell line A549. Treatment with NNN also caused an elevated degree of 5-hydroxymethylcytidine (5hmdC) in DNA, while the methylation degree (i.e. 5-methylcytidine; 5mdC) stayed constant. According to our data, NNN could, via yet unknown mechanisms, play a role in the formation of N3mT as well as 5hmdC. In this study we have developed a new sensitive analytical method using CE-LIF for the simultaneous detection of the three DNA modifications, 5mdC, 5hmdC and N3mT. This article is protected by copyright. All rights reserved.

AB - Methylating substances alter DNA by forming N3-methylthymidine (N3mT), a mutagenic base modification. To develop a sensitive analytical method for the detection of N3mT in DNA based on capillary electrophoresis with laser-induced fluorescence detection (CE-LIF), we synthesized the N3mT-3'-phosphate as a chemical standard. The limit of detection was 1.9 amol of N3mT, which corresponds to one molecule of N3mT per 1,000 normal nucleotides or 0.1%. With this method, we demonstrated that the carcinogenic nitrosamine N'-nitrosonornicotine (NNN) induced N3mT in the human lung cancer cell line A549. Treatment with NNN also caused an elevated degree of 5-hydroxymethylcytidine (5hmdC) in DNA, while the methylation degree (i.e. 5-methylcytidine; 5mdC) stayed constant. According to our data, NNN could, via yet unknown mechanisms, play a role in the formation of N3mT as well as 5hmdC. In this study we have developed a new sensitive analytical method using CE-LIF for the simultaneous detection of the three DNA modifications, 5mdC, 5hmdC and N3mT. This article is protected by copyright. All rights reserved.

KW - 5-hydroxymethylcytosine

KW - Capillary electrophoresis with laser-induced fluorescence

KW - DNA methylation

KW - N3-methylthymidine

KW - Nitrosamines

UR - http://www.scopus.com/inward/record.url?scp=85062329710&partnerID=8YFLogxK

U2 - 10.1002/elps.201800495

DO - 10.1002/elps.201800495

M3 - Article

C2 - 30767246

VL - 40

SP - 1535

EP - 1539

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 11

ER -

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