Artesunate Induces Oxidative DNA Damage, Sustained DNA Double-Strand Breaks, and the ATM/ATR Damage Response in Cancer Cells

Abstract

Abstract Artesunate, the active agent from Artemisia annua L. used in the traditional Chinese medicine, is being applied as a first-line drug for malaria treatment, and trials are ongoing that include this drug in cancer therapy. Despite increasing interest in its therapeutic application, the mode of cell killing provoked by artesunate in human cells is unknown. Here, we show that artesunate is a powerful inducer of oxidative DNA damage, giving rise to formamidopyrimidine DNA glycosylase–sensitive sites and the formation of 8-oxoguanine and 1,N6-ethenoadenine. Oxidative DNA damage was induced in LN-229 human glioblastoma cells dose dependently and was paralleled by cell death executed by apoptosis and necrosis, which could be attenuated by radical scavengers such as N-acetyl cysteine. Oxidative DNA damage resulted in DNA double-strand breaks (DSB) as determined by γH2AX foci that colocalized with 53BP1. Upon chronic treatment with artesunate, the level of DSB continuously increased over the treatment period up to a steady-state level, which is in contrast to ionizing radiation that induced a burst of DSB followed by a decline due to their repair. Knockdown of Rad51 by short interfering RNA and inactivation of DNA-PK strongly sensitized glioma cells to artesunate. These data indicate that both homologous recombination and nonhomologous end joining are involved in the repair of artesunate-induced DSB. Artesunate provoked a DNA damage response (DDR) with phosphorylation of ATM, ATR, Chk1, and Chk2. Overall, these data revealed that artesunate induces oxidative DNA lesions and DSB that continuously increase during the treatment period and accumulate until they trigger DDR and finally tumor cell death. Mol Cancer Ther; 10(12); 2224–33. ©2011 AACR.

Autoren

Nicole Berdelle
Teodora Nikolova
Steve Quiros
Thomas Efferth
Bernd Kaina

DOI

10.1158/1535-7163.mct-11-0534

eISSN

1538-8514

ISSN

1535-7163

Ausgabe der Veröffentlichung

12

Zeitschrift

Molecular Cancer Therapeutics

Sprache

en

Online publication date

2011

Paginierung

2224 - 2233

Datum der Veröffentlichung

2011

Status

Published

Herausgeber

American Association for Cancer Research (AACR)

Herausgeber URL

http://dx.doi.org/10.1158/1535-7163.mct-11-0534

Datum der Datenerfassung

2022

Titel

Artesunate Induces Oxidative DNA Damage, Sustained DNA Double-Strand Breaks, and the ATM/ATR Damage Response in Cancer Cells

Ausgabe der Zeitschrift

10

Data source: Crossref

Abstract

Artesunate, the active agent from Artemisia annua L. used in the traditional Chinese medicine, is being applied as a first-line drug for malaria treatment, and trials are ongoing that include this drug in cancer therapy. Despite increasing interest in its therapeutic application, the mode of cell killing provoked by artesunate in human cells is unknown. Here, we show that artesunate is a powerful inducer of oxidative DNA damage, giving rise to formamidopyrimidine DNA glycosylase-sensitive sites and the formation of 8-oxoguanine and 1,N6-ethenoadenine. Oxidative DNA damage was induced in LN-229 human glioblastoma cells dose dependently and was paralleled by cell death executed by apoptosis and necrosis, which could be attenuated by radical scavengers such as N-acetyl cysteine. Oxidative DNA damage resulted in DNA double-strand breaks (DSB) as determined by γH2AX foci that colocalized with 53BP1. Upon chronic treatment with artesunate, the level of DSB continuously increased over the treatment period up to a steady-state level, which is in contrast to ionizing radiation that induced a burst of DSB followed by a decline due to their repair. Knockdown of Rad51 by short interfering RNA and inactivation of DNA-PK strongly sensitized glioma cells to artesunate. These data indicate that both homologous recombination and nonhomologous end joining are involved in the repair of artesunate-induced DSB. Artesunate provoked a DNA damage response (DDR) with phosphorylation of ATM, ATR, Chk1, and Chk2. Overall, these data revealed that artesunate induces oxidative DNA lesions and DSB that continuously increase during the treatment period and accumulate until they trigger DDR and finally tumor cell death.

Addresses

Institute of Toxicology, University Medical Center, Johannes Gutenberg University, Mainz, Germany.

Autoren

Nicole Berdelle
Teodora Nikolova
Steve Quiros
Thomas Efferth
Thomas Efferth
Bernd Kaina

DOI

10.1158/1535-7163.mct-11-0534

eISSN

1538-8514

Externe Identifier

PubMed Identifier: 21998290

Open access

false

ISSN

1535-7163

Ausgabe der Veröffentlichung

12

Zeitschrift

Molecular cancer therapeutics

Schlüsselwörter

Tumor Cells, Cultured
Humans
Neoplasms
DNA Damage
Artemisinins
Cell Cycle Proteins
DNA-Binding Proteins
Tumor Suppressor Proteins
Antineoplastic Agents, Phytogenic
Drug Evaluation, Preclinical
Apoptosis
DNA Repair
Up-Regulation
Oxidative Stress
Models, Biological
DNA Breaks, Double-Stranded
Ataxia Telangiectasia Mutated Proteins
Artesunate
Protein Serine-Threonine Kinases

Sprache

eng

Medium

Print-Electronic

Online publication date

2011

Paginierung

2224 - 2233

Datum der Veröffentlichung

2011

Status

Published

Datum der Datenerfassung

2011

Titel

Artesunate induces oxidative DNA damage, sustained DNA double-strand breaks, and the ATM/ATR damage response in cancer cells.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

10

Data source: Europe PubMed Central

Abstract

Artesunate, the active agent from Artemisia annua L. used in the traditional Chinese medicine, is being applied as a first-line drug for malaria treatment, and trials are ongoing that include this drug in cancer therapy. Despite increasing interest in its therapeutic application, the mode of cell killing provoked by artesunate in human cells is unknown. Here, we show that artesunate is a powerful inducer of oxidative DNA damage, giving rise to formamidopyrimidine DNA glycosylase-sensitive sites and the formation of 8-oxoguanine and 1,N6-ethenoadenine. Oxidative DNA damage was induced in LN-229 human glioblastoma cells dose dependently and was paralleled by cell death executed by apoptosis and necrosis, which could be attenuated by radical scavengers such as N-acetyl cysteine. Oxidative DNA damage resulted in DNA double-strand breaks (DSB) as determined by γH2AX foci that colocalized with 53BP1. Upon chronic treatment with artesunate, the level of DSB continuously increased over the treatment period up to a steady-state level, which is in contrast to ionizing radiation that induced a burst of DSB followed by a decline due to their repair. Knockdown of Rad51 by short interfering RNA and inactivation of DNA-PK strongly sensitized glioma cells to artesunate. These data indicate that both homologous recombination and nonhomologous end joining are involved in the repair of artesunate-induced DSB. Artesunate provoked a DNA damage response (DDR) with phosphorylation of ATM, ATR, Chk1, and Chk2. Overall, these data revealed that artesunate induces oxidative DNA lesions and DSB that continuously increase during the treatment period and accumulate until they trigger DDR and finally tumor cell death.

Autoren

Nicole Berdelle
Teodora Nikolova
Steve Quiros
Thomas Efferth
Bernd Kaina

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/21998290

DOI

10.1158/1535-7163.MCT-11-0534

eISSN

1538-8514

Ausgabe der Veröffentlichung

12

Zeitschrift

Mol Cancer Ther

Schlüsselwörter

Antineoplastic Agents, Phytogenic
Apoptosis
Artemisinins
Artesunate
Ataxia Telangiectasia Mutated Proteins
Cell Cycle Proteins
DNA Breaks, Double-Stranded
DNA Damage
DNA Repair
DNA-Binding Proteins
Drug Evaluation, Preclinical
Humans
Models, Biological
Neoplasms
Oxidative Stress
Protein Serine-Threonine Kinases
Tumor Cells, Cultured
Tumor Suppressor Proteins
Up-Regulation

Sprache

eng

Country

United States

Paginierung

2224 - 2233

PII

1535-7163.MCT-11-0534

Datum der Veröffentlichung

2011

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2012

Titel

Artesunate induces oxidative DNA damage, sustained DNA double-strand breaks, and the ATM/ATR damage response in cancer cells.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

10

Data source: PubMed

Artesunate Induces Oxidative DNA Damage, Sustained DNA Double-Strand Breaks, and the ATM/ATR Damage Response in Cancer Cells

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