Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1997 Feb 1;321(Pt 3):897–901. doi: 10.1042/bj3210897

Nitric oxide co-operates with hydrogen peroxide in inducing DNA fragmentation and cell lysis in murine lymphoma cells.

J G Filep 1, C Lapierre 1, S Lachance 1, J S Chan 1
PMCID: PMC1218150  PMID: 9032481

Abstract

We examined whether NO and H2O2 could interact in inducing DNA fragmentation and cell death. H2O2 and the NO-releasing compounds sodium nitroprusside (SNP) and S-nitroso-N-acetyl-D,L-penicillamine (SNAP) by themselves elicited lysis of YAC-1 murine lymphoma cells in a concentration-dependent manner. Exposure of the cells to a combination of sublytic concentrations of SNP (0.78 mM) plus H2O2 (7.8 microM) or SNAP (0.18 mM) plus H2O2 (7.8 microM) resulted in cell death which is mediated, in part, through apoptosis. Evidence for this direction is provided by fluorescence microscopic evaluation of the cells, which revealed the presence of changes in nuclear morphology characteristic of apoptosis in 30-40% of lymphoma cells and by the specific pattern of internucleosomal DNA fragmentation detected by gel electrophoresis. The cytotoxic effect of SNP plus H2O2 could be effectively inhibited by either oxyhaemoglobin, which binds NO, or catalase, which eliminates H2O2. Partial protection from SNP-plus-H2O2-induced cell lysis was observed with the poly(ADP-ribose) polymerase inhibitors, nicotinamide and 3-aminobenzamide, parallelling their ability to reverse depletion of cellular NAD+ pools. These results indicate an interaction between NO and H2O2 which leads to a markedly enhanced cytotoxic activity, in part, via induction of apoptosis and suggest that poly(ADP-ribosylation) and subsequent NAD+ depletion mediate, at least in part, this cytotoxic activity.

Full Text

The Full Text of this article is available as a PDF (290.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albina J. E., Cui S., Mateo R. B., Reichner J. S. Nitric oxide-mediated apoptosis in murine peritoneal macrophages. J Immunol. 1993 Jun 1;150(11):5080–5085. [PubMed] [Google Scholar]
  2. Bates J. N., Baker M. T., Guerra R., Jr, Harrison D. G. Nitric oxide generation from nitroprusside by vascular tissue. Evidence that reduction of the nitroprusside anion and cyanide loss are required. Biochem Pharmacol. 1991 Dec 11;42 (Suppl):S157–S165. doi: 10.1016/0006-2952(91)90406-u. [DOI] [PubMed] [Google Scholar]
  3. Beckman J. S., Crow J. P. Pathological implications of nitric oxide, superoxide and peroxynitrite formation. Biochem Soc Trans. 1993 May;21(2):330–334. doi: 10.1042/bst0210330. [DOI] [PubMed] [Google Scholar]
  4. Bernofsky C., Swan M. An improved cycling assay for nicotinamide adenine dinucleotide. Anal Biochem. 1973 Jun;53(2):452–458. doi: 10.1016/0003-2697(73)90094-8. [DOI] [PubMed] [Google Scholar]
  5. Bonfoco E., Krainc D., Ankarcrona M., Nicotera P., Lipton S. A. Apoptosis and necrosis: two distinct events induced, respectively, by mild and intense insults with N-methyl-D-aspartate or nitric oxide/superoxide in cortical cell cultures. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7162–7166. doi: 10.1073/pnas.92.16.7162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brunelli L., Crow J. P., Beckman J. S. The comparative toxicity of nitric oxide and peroxynitrite to Escherichia coli. Arch Biochem Biophys. 1995 Jan 10;316(1):327–334. doi: 10.1006/abbi.1995.1044. [DOI] [PubMed] [Google Scholar]
  7. Buttke T. M., Sandstrom P. A. Oxidative stress as a mediator of apoptosis. Immunol Today. 1994 Jan;15(1):7–10. doi: 10.1016/0167-5699(94)90018-3. [DOI] [PubMed] [Google Scholar]
  8. Duke R. C., Cohen J. J. IL-2 addiction: withdrawal of growth factor activates a suicide program in dependent T cells. Lymphokine Res. 1986 Fall;5(4):289–299. [PubMed] [Google Scholar]
  9. Duke R. C., Witter R. Z., Nash P. B., Young J. D., Ojcius D. M. Cytolysis mediated by ionophores and pore-forming agents: role of intracellular calcium in apoptosis. FASEB J. 1994 Feb;8(2):237–246. doi: 10.1096/fasebj.8.2.8119494. [DOI] [PubMed] [Google Scholar]
  10. Feelisch M., Noack E. A. Correlation between nitric oxide formation during degradation of organic nitrates and activation of guanylate cyclase. Eur J Pharmacol. 1987 Jul 2;139(1):19–30. doi: 10.1016/0014-2999(87)90493-6. [DOI] [PubMed] [Google Scholar]
  11. Hille R., Palmer G., Olson J. S. Chain equivalence in reaction of nitric oxide with hemoglobin. J Biol Chem. 1977 Jan 10;252(1):403–405. [PubMed] [Google Scholar]
  12. Hogg N., Darley-Usmar V. M., Wilson M. T., Moncada S. Production of hydroxyl radicals from the simultaneous generation of superoxide and nitric oxide. Biochem J. 1992 Jan 15;281(Pt 2):419–424. doi: 10.1042/bj2810419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ioannidis I., de Groot H. Cytotoxicity of nitric oxide in Fu5 rat hepatoma cells: evidence for co-operative action with hydrogen peroxide. Biochem J. 1993 Dec 1;296(Pt 2):341–345. doi: 10.1042/bj2960341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kanner J., Harel S., Granit R. Nitric oxide as an antioxidant. Arch Biochem Biophys. 1991 Aug 15;289(1):130–136. doi: 10.1016/0003-9861(91)90452-o. [DOI] [PubMed] [Google Scholar]
  15. Lennon S. V., Martin S. J., Cotter T. G. Dose-dependent induction of apoptosis in human tumour cell lines by widely diverging stimuli. Cell Prolif. 1991 Mar;24(2):203–214. doi: 10.1111/j.1365-2184.1991.tb01150.x. [DOI] [PubMed] [Google Scholar]
  16. Moncada S., Palmer R. M., Higgs E. A. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev. 1991 Jun;43(2):109–142. [PubMed] [Google Scholar]
  17. Nathan C. F. Secretory products of macrophages. J Clin Invest. 1987 Feb;79(2):319–326. doi: 10.1172/JCI112815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nathan C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992 Sep;6(12):3051–3064. [PubMed] [Google Scholar]
  19. Nguyen T., Brunson D., Crespi C. L., Penman B. W., Wishnok J. S., Tannenbaum S. R. DNA damage and mutation in human cells exposed to nitric oxide in vitro. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3030–3034. doi: 10.1073/pnas.89.7.3030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Noronha-Dutra A. A., Epperlein M. M., Woolf N. Reaction of nitric oxide with hydrogen peroxide to produce potentially cytotoxic singlet oxygen as a model for nitric oxide-mediated killing. FEBS Lett. 1993 Apr 19;321(1):59–62. doi: 10.1016/0014-5793(93)80621-z. [DOI] [PubMed] [Google Scholar]
  21. Pacelli R., Wink D. A., Cook J. A., Krishna M. C., DeGraff W., Friedman N., Tsokos M., Samuni A., Mitchell J. B. Nitric oxide potentiates hydrogen peroxide-induced killing of Escherichia coli. J Exp Med. 1995 Nov 1;182(5):1469–1479. doi: 10.1084/jem.182.5.1469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pohajdak B., Gomez J. L., Wilkins J. A., Greenberg A. H. Tumor-activated NK cells trigger monocyte oxidative metabolism. J Immunol. 1984 Nov;133(5):2430–2436. [PubMed] [Google Scholar]
  23. Radons J., Heller B., Bürkle A., Hartmann B., Rodriguez M. L., Kröncke K. D., Burkart V., Kolb H. Nitric oxide toxicity in islet cells involves poly(ADP-ribose) polymerase activation and concomitant NAD+ depletion. Biochem Biophys Res Commun. 1994 Mar 30;199(3):1270–1277. doi: 10.1006/bbrc.1994.1368. [DOI] [PubMed] [Google Scholar]
  24. Schraufstatter I. U., Hyslop P. A., Hinshaw D. B., Spragg R. G., Sklar L. A., Cochrane C. G. Hydrogen peroxide-induced injury of cells and its prevention by inhibitors of poly(ADP-ribose) polymerase. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4908–4912. doi: 10.1073/pnas.83.13.4908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schreck R., Albermann K., Baeuerle P. A. Nuclear factor kappa B: an oxidative stress-responsive transcription factor of eukaryotic cells (a review). Free Radic Res Commun. 1992;17(4):221–237. doi: 10.3109/10715769209079515. [DOI] [PubMed] [Google Scholar]
  26. Schwartzman R. A., Cidlowski J. A. Apoptosis: the biochemistry and molecular biology of programmed cell death. Endocr Rev. 1993 Apr;14(2):133–151. doi: 10.1210/edrv-14-2-133. [DOI] [PubMed] [Google Scholar]
  27. Volk T., Ioannidis I., Hensel M., deGroot H., Kox W. J. Endothelial damage induced by nitric oxide: synergism with reactive oxygen species. Biochem Biophys Res Commun. 1995 Aug 4;213(1):196–203. doi: 10.1006/bbrc.1995.2116. [DOI] [PubMed] [Google Scholar]
  28. Zhang J., Dawson V. L., Dawson T. M., Snyder S. H. Nitric oxide activation of poly(ADP-ribose) synthetase in neurotoxicity. Science. 1994 Feb 4;263(5147):687–689. doi: 10.1126/science.8080500. [DOI] [PubMed] [Google Scholar]
  29. Zhong L. T., Sarafian T., Kane D. J., Charles A. C., Mah S. P., Edwards R. H., Bredesen D. E. bcl-2 inhibits death of central neural cells induced by multiple agents. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4533–4537. doi: 10.1073/pnas.90.10.4533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zingarelli B., O'Connor M., Wong H., Salzman A. L., Szabó C. Peroxynitrite-mediated DNA strand breakage activates poly-adenosine diphosphate ribosyl synthetase and causes cellular energy depletion in macrophages stimulated with bacterial lipopolysaccharide. J Immunol. 1996 Jan 1;156(1):350–358. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES