Poly(ADP-ribosyl)ation is a survival mechanism in cigarette smoke-induced and hydrogen peroxide-mediated cell death

Katalin Kovács, Katalin Erdélyi, Csaba Hegeds, Petra Lakatos, Zsolt Regdon, P. Bai, G. Haskó, Éva Szabó, L. Virag

Research output: Contribution to journalArticle

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Abstract

Cigarette smoking can contribute to the development of many human diseases such as cardiovascular disease, lung cancer, asthma, and chronic obstructive pulmonary disease. Thousands of compounds are present in cigarette smoke, including a large number of reactive oxygen species that can cause DNA damage, leading to the activation of poly(ADP-ribose) polymerase (PARP) enzymes. The PAR polymer is degraded by poly(ADP-ribose) glycohydrolase (PARG). Here we have investigated the effects of cigarette smoke extract (CSE) on A549 human lung epithelial cells. CSE induced DNA damage (comet assay), PAR accumulation (immunofluorescence and immunoblotting), impaired proliferation (clonogenic survival assay and electric cell-substrate impedance sensing measurement), and cell death (MTT reduction, propidium iodide uptake, lactate dehydrogenase release). CSE-induced cell death was also characterized by mitochondrial depolarization but massive translocation of apoptosis-inducing factor could not be observed. To investigate the role of PARylation in CSE-induced oxidative stress, PARP-1- and PARG-silenced A549 cells were used. Silencing of both PARP-1 and PARG sensitized cells to CSE-induced toxicity: PARP-1- and PARG-silenced cell lines exhibited reduced clonogenic survival, displayed a delayed repair of DNA breaks, and showed higher levels of cytotoxicity. CSE triggered the production of mitochondrial superoxide and hydrogen peroxide. Addition of superoxide dismutase increased, whereas catalase abolished, CSE-induced PAR formation. In summary, our data show that the superoxide-hydrogen peroxide-DNA breakage pathway activates the PAR cycle by PARP-1 and PARG, which serves as a survival mechanism in CSE-exposed cells. Our data also raise the possibility that the PARP-1/PARG status of smokers may be an important determinant of the efficiency of DNA repair in their lungs and of their susceptibility to CS-induced carcinogenesis.

Original languageEnglish
Pages (from-to)1680-1688
Number of pages9
JournalFree Radical Biology and Medicine
Volume53
Issue number9
DOIs
Publication statusPublished - Oct 1 2012

Fingerprint

Cell death
Smoke
Tobacco Products
Adenosine Diphosphate
Hydrogen Peroxide
Cell Death
Poly(ADP-ribose) Polymerases
Survival
DNA
Pulmonary diseases
Superoxides
DNA Damage
Assays
Repair
Apoptosis Inducing Factor
Electric batteries
Lung
DNA Breaks
Comet Assay
Propidium

Keywords

  • Cell death
  • Cigarette smoke
  • Free radicals
  • Poly(ADP-ribose) glycohydrolase
  • Poly(ADP-ribose) polymerase
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Poly(ADP-ribosyl)ation is a survival mechanism in cigarette smoke-induced and hydrogen peroxide-mediated cell death. / Kovács, Katalin; Erdélyi, Katalin; Hegeds, Csaba; Lakatos, Petra; Regdon, Zsolt; Bai, P.; Haskó, G.; Szabó, Éva; Virag, L.

In: Free Radical Biology and Medicine, Vol. 53, No. 9, 01.10.2012, p. 1680-1688.

Research output: Contribution to journalArticle

Kovács, Katalin ; Erdélyi, Katalin ; Hegeds, Csaba ; Lakatos, Petra ; Regdon, Zsolt ; Bai, P. ; Haskó, G. ; Szabó, Éva ; Virag, L. / Poly(ADP-ribosyl)ation is a survival mechanism in cigarette smoke-induced and hydrogen peroxide-mediated cell death. In: Free Radical Biology and Medicine. 2012 ; Vol. 53, No. 9. pp. 1680-1688.
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AU - Lakatos, Petra

AU - Regdon, Zsolt

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