Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence.
Krais, Annette LU
; Kliem, Christian
; Arlt, Volker
and Schmeiser, Heinz
(2019)
In Electrophoresis
40(11).
p.1535-1539
- 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;... (More)
- 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 (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/6cd4a680-bda0-41f5-850f-c1b7a9071380
- author
- Krais, Annette
LU
; Kliem, Christian
; Arlt, Volker
and Schmeiser, Heinz
- organization
- publishing date
- 2019-02-15
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Electrophoresis
- volume
- 40
- issue
- 11
- pages
- 1535 - 1539
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:30767246
- scopus:85062329710
- ISSN
- 0173-0835
- DOI
- 10.1002/elps.201800495
- language
- English
- LU publication?
- yes
- id
- 6cd4a680-bda0-41f5-850f-c1b7a9071380
- date added to LUP
- 2019-02-24 22:18:42
- date last changed
- 2022-03-17 21:46:54
@article{6cd4a680-bda0-41f5-850f-c1b7a9071380,
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}},
author = {{Krais, Annette and Kliem, Christian and Arlt, Volker and Schmeiser, Heinz}},
issn = {{0173-0835}},
language = {{eng}},
month = {{02}},
number = {{11}},
pages = {{1535--1539}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Electrophoresis}},
title = {{Determination of genomic N3-methylthymidine in human cancer cells treated with nitrosamines using capillary electrophoresis with laser-induced fluorescence.}},
url = {{http://dx.doi.org/10.1002/elps.201800495}},
doi = {{10.1002/elps.201800495}},
volume = {{40}},
year = {{2019}},
}