Protein oxidative damage and redox imbalance induced by ionising radiation in CHO cells

Donatella Pietraforte, Eleonora Paulicelli, Clarice Patrono, Lucrezia Gambardella, Giuseppe Scorza, Antonella Testa, Paola Fattibene

Research output: Contribution to journalArticle

Abstract

Reactive oxygen species (ROS) are important mediators of the cytotoxicity induced by the direct reaction of ionising radiation (IR) with all critical cellular components, such as proteins, lipids, and nucleic acids. The derived oxidative damage may propagate in exposed tissues in a dose- and spatiotemporal dependent manner to other cell compartments, affecting intracellular signalling, and cell fate. To understand how cell damage is induced, we studied the oxidative events occurring immediately after cell irradiation by analysing the fate of IR-derived ROS, the intracellular oxidative damage, and the modification of redox environment accumulating in Chinese hamster ovary (CHO) within 1 h after cell irradiation (dose range 0-10 Gy). By using the immuno-spin trapping technique (IST), spectrophotometric methods, and electron paramagnetic resonance (EPR) spectroscopy, we showed that IR-derived ROS (i) induced an IST-detectable, antioxidant-inhibitable one-electron oxidation of specific intracellular proteins; (ii) altered the glutathione (GSH) content (which was found to increase below 2 Gy, and decrease at higher doses, leading to a redox imbalance); (iii) decreased glutathione peroxidase and glutaredoxin activity; (iv) modified neither glutathione reductase nor thioredoxin reductase activity; (v) were detected by spin trapping technique, but adduct intensity decreased due to cell competition for ROS; and (vi) induced no EPR-detectable radicals assignable to oxidised cellular components. In conclusion, our results showed that IR generated an early high oxidising potential (protein radical intermediates, redox imbalance, modified redox enzyme activity) in irradiated cells potentially able to propagate the damage and induce oxidative modification of secondary targets.

Original languageEnglish
Pages (from-to)465-479
Number of pages15
JournalFree Radical Research
Volume52
Issue number4
DOIs
Publication statusPublished - Apr 2018

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Ionizing radiation
Cricetulus
Ionizing Radiation
Oxidation-Reduction
Ovary
Reactive Oxygen Species
Spin Trapping
Paramagnetic resonance
Proteins
Cells
Glutaredoxins
Irradiation
Thioredoxin-Disulfide Reductase
Electron Spin Resonance Spectroscopy
Glutathione Reductase
Enzyme activity
Cytotoxicity
Glutathione Peroxidase
Nucleic Acids
Dosimetry

Keywords

  • Animals
  • CHO Cells
  • Cricetulus
  • Glutaredoxins/metabolism
  • Glutathione Peroxidase/metabolism
  • Glutathione Reductase/metabolism
  • Oxidation-Reduction
  • Radiation, Ionizing
  • Reactive Oxygen Species/metabolism

Cite this

Protein oxidative damage and redox imbalance induced by ionising radiation in CHO cells. / Pietraforte, Donatella; Paulicelli, Eleonora; Patrono, Clarice; Gambardella, Lucrezia; Scorza, Giuseppe; Testa, Antonella; Fattibene, Paola.

In: Free Radical Research, Vol. 52, No. 4, 04.2018, p. 465-479.

Research output: Contribution to journalArticle

Pietraforte, Donatella ; Paulicelli, Eleonora ; Patrono, Clarice ; Gambardella, Lucrezia ; Scorza, Giuseppe ; Testa, Antonella ; Fattibene, Paola. / Protein oxidative damage and redox imbalance induced by ionising radiation in CHO cells. In: Free Radical Research. 2018 ; Vol. 52, No. 4. pp. 465-479.
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AU - Pietraforte, Donatella

AU - Paulicelli, Eleonora

AU - Patrono, Clarice

AU - Gambardella, Lucrezia

AU - Scorza, Giuseppe

AU - Testa, Antonella

AU - Fattibene, Paola

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AB - Reactive oxygen species (ROS) are important mediators of the cytotoxicity induced by the direct reaction of ionising radiation (IR) with all critical cellular components, such as proteins, lipids, and nucleic acids. The derived oxidative damage may propagate in exposed tissues in a dose- and spatiotemporal dependent manner to other cell compartments, affecting intracellular signalling, and cell fate. To understand how cell damage is induced, we studied the oxidative events occurring immediately after cell irradiation by analysing the fate of IR-derived ROS, the intracellular oxidative damage, and the modification of redox environment accumulating in Chinese hamster ovary (CHO) within 1 h after cell irradiation (dose range 0-10 Gy). By using the immuno-spin trapping technique (IST), spectrophotometric methods, and electron paramagnetic resonance (EPR) spectroscopy, we showed that IR-derived ROS (i) induced an IST-detectable, antioxidant-inhibitable one-electron oxidation of specific intracellular proteins; (ii) altered the glutathione (GSH) content (which was found to increase below 2 Gy, and decrease at higher doses, leading to a redox imbalance); (iii) decreased glutathione peroxidase and glutaredoxin activity; (iv) modified neither glutathione reductase nor thioredoxin reductase activity; (v) were detected by spin trapping technique, but adduct intensity decreased due to cell competition for ROS; and (vi) induced no EPR-detectable radicals assignable to oxidised cellular components. In conclusion, our results showed that IR generated an early high oxidising potential (protein radical intermediates, redox imbalance, modified redox enzyme activity) in irradiated cells potentially able to propagate the damage and induce oxidative modification of secondary targets.

KW - Animals

KW - CHO Cells

KW - Cricetulus

KW - Glutaredoxins/metabolism

KW - Glutathione Peroxidase/metabolism

KW - Glutathione Reductase/metabolism

KW - Oxidation-Reduction

KW - Radiation, Ionizing

KW - Reactive Oxygen Species/metabolism

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SN - 1071-5762

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