Elaboration of cellular DNA breaks by hydroperoxides
- 31 December 1991
- journal article
- Published by Elsevier BV in Free Radical Biology & Medicine
- Vol. 11 (6), 563-572
- https://doi.org/10.1016/0891-5849(91)90137-r
Abstract
Cellular damage produced by ionizing radiation and peroxides, hydrogen peroxide (HOOH) and the organic peroxides tert-butyl (tBuOOH) or cumene hydroperoxide (CuOOH) were compared. DNA breaks, toxicity, malondialdehyde production, and the rate of peroxide disappearance were measured in a human adenocarcinoma cell line (A549). The alkaline and neutral filter elution assays were used to quantitate the kinetics of single and double strand break formation and repair (SSB and DSB), respectively. Peroxides, at 0.01–1.0 mM, produce multiphasic dose response curves for both toxicity and DNA SSBs. Radiation, 1–6 Gy, produced a shouldered survival curve, and both DNA SSB and DSBs produced in cells x-rayed on ice were nearly linear with dose. The peroxides produced more SSBs than radiation at equitoxic doses. X-ray induced DNA single strand breaks were rejoined rapidly by cells at 37°C with approximately 80% of initial damage repaired in 20 min. Peroxide induced SSBs were maximal after 15 min at 37° C. Rejoining proceeded thereafter, but at a rate less than for x-ray induced strand breaks. Significant DNA DSBs could not be achieved by peroxides even at concentrations 50-fold higher than required to produce SSBs. HOOH treatment of DNA on filters following cells lysis and proteolysis produced SSBs. CuOOH and tBuOOH produced no SSBs in lysed cell DNA. None of the peroxides produced DSBs when incubated with lysed cell DNA. Malondialdehyde was released from cells incubated with organic hydroperoxides, but not HOOH, nor up to 40 Gy of x-rays. HOOH was metabolized three times faster than the organic peroxides. The overall results demonstrate the necessity for a metabolically active cell environment to elaborate maximal DNA strand breaks and cell death at hydroperoxide concentrations of 10−4 of greater, but prevent strand breaks and stimulate cell growth at 10−5M.Keywords
This publication has 28 references indexed in Scilit:
- Selenium nutrition in human lung cells: Response to radiation and peroxidesFree Radical Biology & Medicine, 1990
- Diamide induced shift in protein and glutathione thiol: disulfide status delays DNA rejoining after X-irradiation of human cancer cellsBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1990
- Hydrogen peroxide insult in cultured mammalian cells: relationships between DNA single-strand breakage, poly(ADP-ribose) metabolism and cell killingBiochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1989
- DNA Damage and Oxygen Radical ToxicityScience, 1988
- Toxic DNA Damage by Hydrogen Peroxide Through the Fenton Reaction in Vivo and in VitroScience, 1988
- The Level of Induced DNA Double-strand Breakage Correlates with Cell Killing after X-irradiationInternational Journal of Radiation Biology, 1985
- In vivo formation of single-strand breaks in DNA by hydrogen peroxide is mediated by the Haber-Weiss reactionBiochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1984
- Selenium-dependent and non-selenium-dependent glutathione peroxidases in human tissue extractsBiochimica et Biophysica Acta (BBA) - General Subjects, 1983
- DNA Double Strand Breaks in Ehrlich Ascites Tumour Cells at Low Doses of X-rays. II. Can Cell Death be Attributed to Double Strand Breaks?International Journal of Radiation Biology and Related Studies in Physics, Chemistry and Medicine, 1982
- Comparison of the effects of hydrogen peroxide and X-ray irradiation on toxicity, mutation, and DNA damage/repair in mammalian cells (V-79)Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1981