Apoptosis induced by extracellular glutathione is mediated by H2O2 production and DNA damage

Paola Perego, Laura Gatti, Nives Carenini, Laura Dal Bo, Franco Zunino

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The biochemical basis of the anti-proliferative effect of exogenous glutathione was investigated in A2780 ovarian carcinoma cells. Previous observations have implicated γ-glutamyl transpeptidase-mediated pro-oxidant reactions as a primary mechanism of the extracellular effects of glutathione. In 2 cell lines (A2780 and IGROV-1), glutathione led to H2O2 production, but only A2780 cells, characterized by low expression of detoxification enzymes, were sensitive to the thiol compound. In A2780 cells, glutathione exposure resulted in DNA single-strand break formation, as measured by alkaline elution. Glutathione-induced DNA damage generated significant levels of apoptosis in A2780 cells, but not in IGROV-I cells. The capability of glutathione to induce apoptosis was associated with cleavage of poly(ADP- ribose)polymerase and with generation of a low-molecular-weight form of the pro-apoptotic protein bax. In A2780 cells, glutathione exposure was followed by p21 and Bax induction and p53 up-regulation, as expected for genotoxic stress. Consistently, analysis of cell-cycle perturbations demonstrated the occurrence of G2M accumulation after exposure to glutathione, similar to what was observed for H2O2. Taken together, these results indicate that the cytotoxic effect of extracellular glutathione, related to membrane metabolism, is mediated by production of H2O2 leading to DNA damage and a cellular response involving p53. These findings might also provide insights into the cellular and molecular determinants of chemosensitivity to DNA damaging agents, as oxidative stress is implicated in p53-dependent apoptosis. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish
Pages (from-to)343-348
Number of pages6
JournalInternational Journal of Cancer
Volume87
Issue number3
DOIs
Publication statusPublished - Aug 1 2000

Fingerprint

DNA Damage
Glutathione
Apoptosis
Single-Stranded DNA Breaks
Apoptosis Regulatory Proteins
Poly(ADP-ribose) Polymerases
gamma-Glutamyltransferase
Sulfhydryl Compounds
Reactive Oxygen Species
Cell Cycle
Oxidative Stress
Up-Regulation
Molecular Weight
Carcinoma
Cell Line
Membranes
DNA
Enzymes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Apoptosis induced by extracellular glutathione is mediated by H2O2 production and DNA damage. / Perego, Paola; Gatti, Laura; Carenini, Nives; Dal Bo, Laura; Zunino, Franco.

In: International Journal of Cancer, Vol. 87, No. 3, 01.08.2000, p. 343-348.

Research output: Contribution to journalArticle

@article{9d6f2a6de91c4625b0789aabd3f40802,
title = "Apoptosis induced by extracellular glutathione is mediated by H2O2 production and DNA damage",
abstract = "The biochemical basis of the anti-proliferative effect of exogenous glutathione was investigated in A2780 ovarian carcinoma cells. Previous observations have implicated γ-glutamyl transpeptidase-mediated pro-oxidant reactions as a primary mechanism of the extracellular effects of glutathione. In 2 cell lines (A2780 and IGROV-1), glutathione led to H2O2 production, but only A2780 cells, characterized by low expression of detoxification enzymes, were sensitive to the thiol compound. In A2780 cells, glutathione exposure resulted in DNA single-strand break formation, as measured by alkaline elution. Glutathione-induced DNA damage generated significant levels of apoptosis in A2780 cells, but not in IGROV-I cells. The capability of glutathione to induce apoptosis was associated with cleavage of poly(ADP- ribose)polymerase and with generation of a low-molecular-weight form of the pro-apoptotic protein bax. In A2780 cells, glutathione exposure was followed by p21 and Bax induction and p53 up-regulation, as expected for genotoxic stress. Consistently, analysis of cell-cycle perturbations demonstrated the occurrence of G2M accumulation after exposure to glutathione, similar to what was observed for H2O2. Taken together, these results indicate that the cytotoxic effect of extracellular glutathione, related to membrane metabolism, is mediated by production of H2O2 leading to DNA damage and a cellular response involving p53. These findings might also provide insights into the cellular and molecular determinants of chemosensitivity to DNA damaging agents, as oxidative stress is implicated in p53-dependent apoptosis. (C) 2000 Wiley-Liss, Inc.",
author = "Paola Perego and Laura Gatti and Nives Carenini and {Dal Bo}, Laura and Franco Zunino",
year = "2000",
month = "8",
day = "1",
doi = "10.1002/1097-0215(20000801)87:3<343::AID-IJC6>3.0.CO;2-8",
language = "English",
volume = "87",
pages = "343--348",
journal = "International Journal of Cancer",
issn = "0020-7136",
publisher = "Wiley-Liss Inc.",
number = "3",

}

TY - JOUR

T1 - Apoptosis induced by extracellular glutathione is mediated by H2O2 production and DNA damage

AU - Perego, Paola

AU - Gatti, Laura

AU - Carenini, Nives

AU - Dal Bo, Laura

AU - Zunino, Franco

PY - 2000/8/1

Y1 - 2000/8/1

N2 - The biochemical basis of the anti-proliferative effect of exogenous glutathione was investigated in A2780 ovarian carcinoma cells. Previous observations have implicated γ-glutamyl transpeptidase-mediated pro-oxidant reactions as a primary mechanism of the extracellular effects of glutathione. In 2 cell lines (A2780 and IGROV-1), glutathione led to H2O2 production, but only A2780 cells, characterized by low expression of detoxification enzymes, were sensitive to the thiol compound. In A2780 cells, glutathione exposure resulted in DNA single-strand break formation, as measured by alkaline elution. Glutathione-induced DNA damage generated significant levels of apoptosis in A2780 cells, but not in IGROV-I cells. The capability of glutathione to induce apoptosis was associated with cleavage of poly(ADP- ribose)polymerase and with generation of a low-molecular-weight form of the pro-apoptotic protein bax. In A2780 cells, glutathione exposure was followed by p21 and Bax induction and p53 up-regulation, as expected for genotoxic stress. Consistently, analysis of cell-cycle perturbations demonstrated the occurrence of G2M accumulation after exposure to glutathione, similar to what was observed for H2O2. Taken together, these results indicate that the cytotoxic effect of extracellular glutathione, related to membrane metabolism, is mediated by production of H2O2 leading to DNA damage and a cellular response involving p53. These findings might also provide insights into the cellular and molecular determinants of chemosensitivity to DNA damaging agents, as oxidative stress is implicated in p53-dependent apoptosis. (C) 2000 Wiley-Liss, Inc.

AB - The biochemical basis of the anti-proliferative effect of exogenous glutathione was investigated in A2780 ovarian carcinoma cells. Previous observations have implicated γ-glutamyl transpeptidase-mediated pro-oxidant reactions as a primary mechanism of the extracellular effects of glutathione. In 2 cell lines (A2780 and IGROV-1), glutathione led to H2O2 production, but only A2780 cells, characterized by low expression of detoxification enzymes, were sensitive to the thiol compound. In A2780 cells, glutathione exposure resulted in DNA single-strand break formation, as measured by alkaline elution. Glutathione-induced DNA damage generated significant levels of apoptosis in A2780 cells, but not in IGROV-I cells. The capability of glutathione to induce apoptosis was associated with cleavage of poly(ADP- ribose)polymerase and with generation of a low-molecular-weight form of the pro-apoptotic protein bax. In A2780 cells, glutathione exposure was followed by p21 and Bax induction and p53 up-regulation, as expected for genotoxic stress. Consistently, analysis of cell-cycle perturbations demonstrated the occurrence of G2M accumulation after exposure to glutathione, similar to what was observed for H2O2. Taken together, these results indicate that the cytotoxic effect of extracellular glutathione, related to membrane metabolism, is mediated by production of H2O2 leading to DNA damage and a cellular response involving p53. These findings might also provide insights into the cellular and molecular determinants of chemosensitivity to DNA damaging agents, as oxidative stress is implicated in p53-dependent apoptosis. (C) 2000 Wiley-Liss, Inc.

UR - http://www.scopus.com/inward/record.url?scp=0034255749&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034255749&partnerID=8YFLogxK

U2 - 10.1002/1097-0215(20000801)87:3<343::AID-IJC6>3.0.CO;2-8

DO - 10.1002/1097-0215(20000801)87:3<343::AID-IJC6>3.0.CO;2-8

M3 - Article

C2 - 10897038

AN - SCOPUS:0034255749

VL - 87

SP - 343

EP - 348

JO - International Journal of Cancer

JF - International Journal of Cancer

SN - 0020-7136

IS - 3

ER -