Ascorbate as a "redox sensor" and protector against irradiation-induced oxidative stress in 32D CL 3 hematopoietic cells and subclones overexpressing human manganese superoxide dismutase

Michael W. Epperly, Anatoli N. Osipov, Ian Martin, Kazuaki K. Kawai, Grigory G. Borisenko, Yulia Y. Tyurina, Mia Jefferson, Michael Bernarding, Joel S. Greenberger, Valerian E. Kagan

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Purpose: To determine whether increased expression of manganese superoxide dismutase (MnSOD) protects cells from irradiation by preventing the production of reactive oxygen species (ROS), a new approach to detecting free radical intermediates using ascorbate as an endogenous spin trap was used. Materials and Methods: Cells from the 32D cl 3 hematopoietic cell line or a subclone overexpressing MnSOD (2C6) were incubated with dehydroascorbate for 30 min and irradiated to doses from 0 to 50 Gy. Radical intermediates reacting with spin traps or ascorbate were measured by electron spin resonance spectroscopy. Results were compared to irradiation-induced changes in thiol levels, irradiation survival curves, and accumulation of 8-OHdG as a measurement of DNA oxidative damage. Results: Manganese superoxide dismutase-overexpressing 2C6 cells maintained higher levels of ascorbate (5.4 ± 0.5 and 2.6 ± 0.5 nmol/106 cells, respectively) and thiols (14.0 ± 0.1 and 11.1 ± 0.2 nmol/106 cells) compared to 32D cl 3 parent cells. Cells overexpressing MnSOD produced lower levels of ROS than did the parental 32D cl 3 cells, as evidenced by lower expenditure of ascorbate and GSH after irradiation. Increased ascorbate levels protected both 32D cl 3 and 2C6 cells from irradiation killing, as demonstrated by an increased shoulder on survival curves and decreased DNA 8-OHdG accumulation. Conclusions: Manganese superoxide dismutase overexpression protects 2C6 cells from irradiation damage by scavenging ROS that readily interact with major endogenous antioxidants - ascorbate and GSH - in nontransfected hematopoietic 32D cl 3 cells.

Original languageEnglish (US)
Pages (from-to)851-861
Number of pages11
JournalInternational Journal of Radiation Oncology Biology Physics
Volume58
Issue number3
DOIs
StatePublished - Mar 2004
Externally publishedYes

Fingerprint

protectors
inorganic peroxides
Superoxide Dismutase
Oxidation-Reduction
manganese
Oxidative Stress
irradiation
sensors
cells
Reactive Oxygen Species
Sulfhydryl Compounds
thiols
oxygen
deoxyribonucleic acid
traps
damage
Survival
scavenging
Electron Spin Resonance Spectroscopy
antioxidants

Keywords

  • Ascorbate
  • MnSOD
  • Reactive oxygen species
  • Spin trap

ASJC Scopus subject areas

  • Oncology
  • Radiology Nuclear Medicine and imaging
  • Radiation

Cite this

Ascorbate as a "redox sensor" and protector against irradiation-induced oxidative stress in 32D CL 3 hematopoietic cells and subclones overexpressing human manganese superoxide dismutase. / Epperly, Michael W.; Osipov, Anatoli N.; Martin, Ian; Kawai, Kazuaki K.; Borisenko, Grigory G.; Tyurina, Yulia Y.; Jefferson, Mia; Bernarding, Michael; Greenberger, Joel S.; Kagan, Valerian E.

In: International Journal of Radiation Oncology Biology Physics, Vol. 58, No. 3, 03.2004, p. 851-861.

Research output: Contribution to journalArticle

Epperly, Michael W. ; Osipov, Anatoli N. ; Martin, Ian ; Kawai, Kazuaki K. ; Borisenko, Grigory G. ; Tyurina, Yulia Y. ; Jefferson, Mia ; Bernarding, Michael ; Greenberger, Joel S. ; Kagan, Valerian E. / Ascorbate as a "redox sensor" and protector against irradiation-induced oxidative stress in 32D CL 3 hematopoietic cells and subclones overexpressing human manganese superoxide dismutase. In: International Journal of Radiation Oncology Biology Physics. 2004 ; Vol. 58, No. 3. pp. 851-861.
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abstract = "Purpose: To determine whether increased expression of manganese superoxide dismutase (MnSOD) protects cells from irradiation by preventing the production of reactive oxygen species (ROS), a new approach to detecting free radical intermediates using ascorbate as an endogenous spin trap was used. Materials and Methods: Cells from the 32D cl 3 hematopoietic cell line or a subclone overexpressing MnSOD (2C6) were incubated with dehydroascorbate for 30 min and irradiated to doses from 0 to 50 Gy. Radical intermediates reacting with spin traps or ascorbate were measured by electron spin resonance spectroscopy. Results were compared to irradiation-induced changes in thiol levels, irradiation survival curves, and accumulation of 8-OHdG as a measurement of DNA oxidative damage. Results: Manganese superoxide dismutase-overexpressing 2C6 cells maintained higher levels of ascorbate (5.4 ± 0.5 and 2.6 ± 0.5 nmol/106 cells, respectively) and thiols (14.0 ± 0.1 and 11.1 ± 0.2 nmol/106 cells) compared to 32D cl 3 parent cells. Cells overexpressing MnSOD produced lower levels of ROS than did the parental 32D cl 3 cells, as evidenced by lower expenditure of ascorbate and GSH after irradiation. Increased ascorbate levels protected both 32D cl 3 and 2C6 cells from irradiation killing, as demonstrated by an increased shoulder on survival curves and decreased DNA 8-OHdG accumulation. Conclusions: Manganese superoxide dismutase overexpression protects 2C6 cells from irradiation damage by scavenging ROS that readily interact with major endogenous antioxidants - ascorbate and GSH - in nontransfected hematopoietic 32D cl 3 cells.",
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T1 - Ascorbate as a "redox sensor" and protector against irradiation-induced oxidative stress in 32D CL 3 hematopoietic cells and subclones overexpressing human manganese superoxide dismutase

AU - Epperly, Michael W.

AU - Osipov, Anatoli N.

AU - Martin, Ian

AU - Kawai, Kazuaki K.

AU - Borisenko, Grigory G.

AU - Tyurina, Yulia Y.

AU - Jefferson, Mia

AU - Bernarding, Michael

AU - Greenberger, Joel S.

AU - Kagan, Valerian E.

PY - 2004/3

Y1 - 2004/3

N2 - Purpose: To determine whether increased expression of manganese superoxide dismutase (MnSOD) protects cells from irradiation by preventing the production of reactive oxygen species (ROS), a new approach to detecting free radical intermediates using ascorbate as an endogenous spin trap was used. Materials and Methods: Cells from the 32D cl 3 hematopoietic cell line or a subclone overexpressing MnSOD (2C6) were incubated with dehydroascorbate for 30 min and irradiated to doses from 0 to 50 Gy. Radical intermediates reacting with spin traps or ascorbate were measured by electron spin resonance spectroscopy. Results were compared to irradiation-induced changes in thiol levels, irradiation survival curves, and accumulation of 8-OHdG as a measurement of DNA oxidative damage. Results: Manganese superoxide dismutase-overexpressing 2C6 cells maintained higher levels of ascorbate (5.4 ± 0.5 and 2.6 ± 0.5 nmol/106 cells, respectively) and thiols (14.0 ± 0.1 and 11.1 ± 0.2 nmol/106 cells) compared to 32D cl 3 parent cells. Cells overexpressing MnSOD produced lower levels of ROS than did the parental 32D cl 3 cells, as evidenced by lower expenditure of ascorbate and GSH after irradiation. Increased ascorbate levels protected both 32D cl 3 and 2C6 cells from irradiation killing, as demonstrated by an increased shoulder on survival curves and decreased DNA 8-OHdG accumulation. Conclusions: Manganese superoxide dismutase overexpression protects 2C6 cells from irradiation damage by scavenging ROS that readily interact with major endogenous antioxidants - ascorbate and GSH - in nontransfected hematopoietic 32D cl 3 cells.

AB - Purpose: To determine whether increased expression of manganese superoxide dismutase (MnSOD) protects cells from irradiation by preventing the production of reactive oxygen species (ROS), a new approach to detecting free radical intermediates using ascorbate as an endogenous spin trap was used. Materials and Methods: Cells from the 32D cl 3 hematopoietic cell line or a subclone overexpressing MnSOD (2C6) were incubated with dehydroascorbate for 30 min and irradiated to doses from 0 to 50 Gy. Radical intermediates reacting with spin traps or ascorbate were measured by electron spin resonance spectroscopy. Results were compared to irradiation-induced changes in thiol levels, irradiation survival curves, and accumulation of 8-OHdG as a measurement of DNA oxidative damage. Results: Manganese superoxide dismutase-overexpressing 2C6 cells maintained higher levels of ascorbate (5.4 ± 0.5 and 2.6 ± 0.5 nmol/106 cells, respectively) and thiols (14.0 ± 0.1 and 11.1 ± 0.2 nmol/106 cells) compared to 32D cl 3 parent cells. Cells overexpressing MnSOD produced lower levels of ROS than did the parental 32D cl 3 cells, as evidenced by lower expenditure of ascorbate and GSH after irradiation. Increased ascorbate levels protected both 32D cl 3 and 2C6 cells from irradiation killing, as demonstrated by an increased shoulder on survival curves and decreased DNA 8-OHdG accumulation. Conclusions: Manganese superoxide dismutase overexpression protects 2C6 cells from irradiation damage by scavenging ROS that readily interact with major endogenous antioxidants - ascorbate and GSH - in nontransfected hematopoietic 32D cl 3 cells.

KW - Ascorbate

KW - MnSOD

KW - Reactive oxygen species

KW - Spin trap

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