Skip to main content
NCBI home page
Environmental Health Perspectives logoLink to Environmental Health Perspectives
. 1997 Sep;105(Suppl 5):1291–1296. doi: 10.1289/ehp.97105s51291

Formation and persistence of 8-oxoguanine in rat lung cells as an important determinant for tumor formation following particle exposure.

P Nehls 1, F Seiler 1, B Rehn 1, R Greferath 1, J Bruch 1
PMCID: PMC1470144  PMID: 9400740

Abstract

Exposure of rats to quartz (or various other particles) can lead to the development of lung tumors. At the moment, the mechanisms involved in particle-induced tumor formation are not clarified. However, it is suggested that inflammation, in conjunction with the production of reactive oxygen species (ROS) and an enhancement of epithelial cell proliferation, may play a key role in the development of lung tumors. ROS induces 8-oxoguanine (8-oxoGua) and other mutagenic DNA oxidation products, which can be converted to mutations in proliferating cells. Mutation formation in cancer-related genes is a critical event with respect to tumor formation. In this study we investigated the effects of quartz (DQ12) and of the nontumorigenic dust corundum on the induction of 8-oxoGua in the DNA of rat lung cells, as well as on cell proliferation and pulmonary inflammation. Wistar rats were exposed by intratracheal instillation to quartz (2.5 mg/rat) or corundum (2.5 mg/rat) suspended in physiological saline; control animals exposed to physiological saline or left untreated. Measurements were carried out 7, 21, and 90 days after the exposures. 8-oxoGua levels were determined in lung tissue sections at the single cell level by immunocytological assay using a rabbit anti-8-oxoGua antibody. After exposure to quartz, 8-oxoGua levels were significantly increased at all time points of investigation. Additionally, we observed inflammation and an enhanced cell proliferation. Exposure to corundum had no adverse effects on the lung; neither increased 8-oxoGua levels nor enhanced cell proliferation or inflammation were detected. These observations support the suggestion that inflammation associated with increased 8-oxoGua levels in lung cells and increased cell proliferation is an important determinant for particle-induced development of lung tumors in the rat.

Full text

PDF
1293

Images in this article

1293Figure 1. A i
on p.1293
1293Figure 1. A ii
on p.1293
1293Figure 1. A iii
on p.1293
1293Figure 1. B
on p.1293
1293Figure 1. B
on p.1293
1293Figure 1. B
on p.1293
1293Figure 1. C
on p.1293
1293Figure 1. C
on p.1293
1293Figure 1. C
on p.1293

Selected References

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

  1. Aggarwal B. B., Kohr W. J., Hass P. E., Moffat B., Spencer S. A., Henzel W. J., Bringman T. S., Nedwin G. E., Goeddel D. V., Harkins R. N. Human tumor necrosis factor. Production, purification, and characterization. J Biol Chem. 1985 Feb 25;260(4):2345–2354. [PubMed] [Google Scholar]
  2. Ames B. N., Gold L. S. Endogenous mutagens and the causes of aging and cancer. Mutat Res. 1991 Sep-Oct;250(1-2):3–16. doi: 10.1016/0027-5107(91)90157-j. [DOI] [PubMed] [Google Scholar]
  3. Anderson D. Antioxidant defences against reactive oxygen species causing genetic and other damage. Mutat Res. 1996 Feb 19;350(1):103–108. doi: 10.1016/0027-5107(95)00096-8. [DOI] [PubMed] [Google Scholar]
  4. Barrett J. C. Mechanisms of multistep carcinogenesis and carcinogen risk assessment. Environ Health Perspect. 1993 Apr;100:9–20. doi: 10.1289/ehp.931009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bessho T., Tano K., Kasai H., Ohtsuka E., Nishimura S. Evidence for two DNA repair enzymes for 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) in human cells. J Biol Chem. 1993 Sep 15;268(26):19416–19421. [PubMed] [Google Scholar]
  6. Bruch J., Gono E., Malkusch W., Rehn B. Improved method for quantitative analysis of lung surfactant phospholipids in bronchoalveolar lavage fluids by high-performance liquid chromatography. Clin Chim Acta. 1994 Dec 16;231(2):193–204. doi: 10.1016/0009-8981(94)90203-8. [DOI] [PubMed] [Google Scholar]
  7. Bruch J., Rehn B., Song H., Gono E., Malkusch W. Toxicological investigations on silicon carbide. 1. Inhalation studies. Br J Ind Med. 1993 Sep;50(9):797–806. doi: 10.1136/oem.50.9.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cerutti P. A. Oxy-radicals and cancer. Lancet. 1994 Sep 24;344(8926):862–863. doi: 10.1016/s0140-6736(94)92832-0. [DOI] [PubMed] [Google Scholar]
  9. Cerutti P., Ghosh R., Oya Y., Amstad P. The role of the cellular antioxidant defense in oxidant carcinogenesis. Environ Health Perspect. 1994 Dec;102 (Suppl 10):123–129. doi: 10.1289/ehp.94102s10123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chao C. C., Park S. H., Aust A. E. Participation of nitric oxide and iron in the oxidation of DNA in asbestos-treated human lung epithelial cells. Arch Biochem Biophys. 1996 Feb 1;326(1):152–157. doi: 10.1006/abbi.1996.0059. [DOI] [PubMed] [Google Scholar]
  11. Clayson D. B., Mehta R., Iverson F. International Commission for Protection Against Environmental Mutagens and Carcinogens. Oxidative DNA damage--the effects of certain genotoxic and operationally non-genotoxic carcinogens. Mutat Res. 1994 Feb;317(1):25–42. doi: 10.1016/0165-1110(94)90010-8. [DOI] [PubMed] [Google Scholar]
  12. Demple B., Harrison L. Repair of oxidative damage to DNA: enzymology and biology. Annu Rev Biochem. 1994;63:915–948. doi: 10.1146/annurev.bi.63.070194.004411. [DOI] [PubMed] [Google Scholar]
  13. Dizdaroglu M. Chemical determination of oxidative DNA damage by gas chromatography-mass spectrometry. Methods Enzymol. 1994;234:3–16. doi: 10.1016/0076-6879(94)34072-2. [DOI] [PubMed] [Google Scholar]
  14. Dreher D., Junod A. F. Role of oxygen free radicals in cancer development. Eur J Cancer. 1996 Jan;32A(1):30–38. doi: 10.1016/0959-8049(95)00531-5. [DOI] [PubMed] [Google Scholar]
  15. Driscoll K. E., Carter J. M., Howard B. W., Hassenbein D. G., Pepelko W., Baggs R. B., Oberdörster G. Pulmonary inflammatory, chemokine, and mutagenic responses in rats after subchronic inhalation of carbon black. Toxicol Appl Pharmacol. 1996 Feb;136(2):372–380. doi: 10.1006/taap.1996.0045. [DOI] [PubMed] [Google Scholar]
  16. Driscoll K. E., Deyo L. C., Howard B. W., Poynter J., Carter J. M. Characterizing mutagenesis in the hprt gene of rat alveolar epithelial cells. Exp Lung Res. 1995 Nov-Dec;21(6):941–956. doi: 10.3109/01902149509031772. [DOI] [PubMed] [Google Scholar]
  17. ERLANGER B. F., BEISER S. M. ANTIBODIES SPECIFIC FOR RIBONUCLEOSIDES AND RIBONUCLEOTIDES AND THEIR REACTION WITH DNA. Proc Natl Acad Sci U S A. 1964 Jul;52:68–74. doi: 10.1073/pnas.52.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  19. Fearon E. R., Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990 Jun 1;61(5):759–767. doi: 10.1016/0092-8674(90)90186-i. [DOI] [PubMed] [Google Scholar]
  20. Feig D. I., Reid T. M., Loeb L. A. Reactive oxygen species in tumorigenesis. Cancer Res. 1994 Apr 1;54(7 Suppl):1890s–1894s. [PubMed] [Google Scholar]
  21. Floyd R. A. The role of 8-hydroxyguanine in carcinogenesis. Carcinogenesis. 1990 Sep;11(9):1447–1450. doi: 10.1093/carcin/11.9.1447. [DOI] [PubMed] [Google Scholar]
  22. Friemann J., Pott F., Wilk K. B., Ahrens O., Rosenbruch M., Hilscher W., Schlipköter H. W. Pulmonary alveolar proteinosis in rats after administration of quartz: its possible role in morphogenesis of lung cancer. J Cancer Res Clin Oncol. 1994;120(6):348–353. doi: 10.1007/BF01247459. [DOI] [PubMed] [Google Scholar]
  23. Gallagher J., Heinrich U., George M., Hendee L., Phillips D. H., Lewtas J. Formation of DNA adducts in rat lung following chronic inhalation of diesel emissions, carbon black and titanium dioxide particles. Carcinogenesis. 1994 Jul;15(7):1291–1299. doi: 10.1093/carcin/15.7.1291. [DOI] [PubMed] [Google Scholar]
  24. Halliwell B., Dizdaroglu M. The measurement of oxidative damage to DNA by HPLC and GC/MS techniques. Free Radic Res Commun. 1992;16(2):75–87. doi: 10.3109/10715769209049161. [DOI] [PubMed] [Google Scholar]
  25. Hatahet Z., Kow Y. W., Purmal A. A., Cunningham R. P., Wallace S. S. New substrates for old enzymes. 5-Hydroxy-2'-deoxycytidine and 5-hydroxy-2'-deoxyuridine are substrates for Escherichia coli endonuclease III and formamidopyrimidine DNA N-glycosylase, while 5-hydroxy-2'-deoxyuridine is a substrate for uracil DNA N-glycosylase. J Biol Chem. 1994 Jul 22;269(29):18814–18820. [PubMed] [Google Scholar]
  26. Jackson J. H. Potential molecular mechanisms of oxidant-induced carcinogenesis. Environ Health Perspect. 1994 Dec;102 (Suppl 10):155–157. doi: 10.1289/ehp.94102s10155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jaruga P., Dizdaroglu M. Repair of products of oxidative DNA base damage in human cells. Nucleic Acids Res. 1996 Apr 15;24(8):1389–1394. doi: 10.1093/nar/24.8.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kamiya H., Miura H., Murata-Kamiya N., Ishikawa H., Sakaguchi T., Inoue H., Sasaki T., Masutani C., Hanaoka F., Nishimura S. 8-Hydroxyadenine (7,8-dihydro-8-oxoadenine) induces misincorporation in in vitro DNA synthesis and mutations in NIH 3T3 cells. Nucleic Acids Res. 1995 Aug 11;23(15):2893–2899. doi: 10.1093/nar/23.15.2893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kasai H., Nishimura S. Hydroxylation of deoxyguanosine at the C-8 position by ascorbic acid and other reducing agents. Nucleic Acids Res. 1984 Feb 24;12(4):2137–2145. doi: 10.1093/nar/12.4.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  31. Mauderly J. L., Jones R. K., Griffith W. C., Henderson R. F., McClellan R. O. Diesel exhaust is a pulmonary carcinogen in rats exposed chronically by inhalation. Fundam Appl Toxicol. 1987 Aug;9(2):208–221. doi: 10.1016/0272-0590(87)90044-3. [DOI] [PubMed] [Google Scholar]
  32. Moyer V. D., Cistulli C. A., Vaslet C. A., Kane A. B. Oxygen radicals and asbestos carcinogenesis. Environ Health Perspect. 1994 Dec;102 (Suppl 10):131–136. doi: 10.1289/ehp.94102s10131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Muhle H., Takenaka S., Mohr U., Dasenbrock C., Mermelstein R. Lung tumor induction upon long-term low-level inhalation of crystalline silica. Am J Ind Med. 1989;15(3):343–346. doi: 10.1002/ajim.4700150309. [DOI] [PubMed] [Google Scholar]
  34. Müller R. Calculation of average antibody affinity in anti-hapten sera from data obtained by competitive radioimmunoassay. J Immunol Methods. 1980;34(4):345–352. doi: 10.1016/0022-1759(80)90107-6. [DOI] [PubMed] [Google Scholar]
  35. Nehls P., Adamkiewicz J., Rajewsky M. F. Immuno-slot-blot: a highly sensitive immunoassay for the quantitation of carcinogen-modified nucleosides in DNA. J Cancer Res Clin Oncol. 1984;108(1):23–29. doi: 10.1007/BF00390969. [DOI] [PubMed] [Google Scholar]
  36. Nehls P., Rajewsky M. F. Differential formation of O6-ethylguanine in the DNA of rat brain chromatin fibers of different folding levels exposed to N-ethyl-N-nitrosourea in vitro. Cancer Res. 1985 Mar;45(3):1378–1383. [PubMed] [Google Scholar]
  37. Nehls P., Rajewsky M. F. Monoclonal antibody-based immunoassay for the determination of cellular enzymatic activity for repair of specific carcinogen-DNA adducts (O6-alkylguanine). Carcinogenesis. 1990 Jan;11(1):81–87. doi: 10.1093/carcin/11.1.81. [DOI] [PubMed] [Google Scholar]
  38. Nehls P., Rajewsky M. F., Spiess E., Werner D. Highly sensitive sites for guanine-O6 ethylation in rat brain DNA exposed to N-ethyl-N-nitrosourea in vivo. EMBO J. 1984 Feb;3(2):327–332. doi: 10.1002/j.1460-2075.1984.tb01806.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Purmal A. A., Kow Y. W., Wallace S. S. Major oxidative products of cytosine, 5-hydroxycytosine and 5-hydroxyuracil, exhibit sequence context-dependent mispairing in vitro. Nucleic Acids Res. 1994 Jan 11;22(1):72–78. doi: 10.1093/nar/22.1.72. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Quinlan T. R., Marsh J. P., Janssen Y. M., Borm P. A., Mossman B. T. Oxygen radicals and asbestos-mediated disease. Environ Health Perspect. 1994 Dec;102 (Suppl 10):107–110. doi: 10.1289/ehp.94102s10107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Seiler F., Kirstein U., Eberle G., Hochleitner K., Rajewsky M. F. Quantification of specific DNA O-alkylation products in individual cells by monoclonal antibodies and digital imaging of intensified nuclear fluorescence. Carcinogenesis. 1993 Sep;14(9):1907–1913. doi: 10.1093/carcin/14.9.1907. [DOI] [PubMed] [Google Scholar]
  42. Shi X., Mao Y., Daniel L. N., Saffiotti U., Dalal N. S., Vallyathan V. Silica radical-induced DNA damage and lipid peroxidation. Environ Health Perspect. 1994 Dec;102 (Suppl 10):149–154. doi: 10.1289/ehp.94102s10149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sugimura T. Multistep carcinogenesis: a 1992 perspective. Science. 1992 Oct 23;258(5082):603–607. doi: 10.1126/science.1411570. [DOI] [PubMed] [Google Scholar]
  44. Thomale J., Huh N. H., Nehls P., Eberle G., Rajewsky M. F. Repair of O6-ethylguanine in DNA protects rat 208F cells from tumorigenic conversion by N-ethyl-N-nitrosourea. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9883–9887. doi: 10.1073/pnas.87.24.9883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Vallyathan V. Generation of oxygen radicals by minerals and its correlation to cytotoxicity. Environ Health Perspect. 1994 Dec;102 (Suppl 10):111–115. doi: 10.1289/ehp.94102s10111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yamano Y., Kagawa J., Hanaoka T., Takahashi T., Kasai H., Tsugane S., Watanabe S. Oxidative DNA damage induced by silica in vivo. Environ Res. 1995 May;69(2):102–107. doi: 10.1006/enrs.1995.1031. [DOI] [PubMed] [Google Scholar]

Articles from Environmental Health Perspectives are provided here courtesy of National Institute of Environmental Health Sciences

RESOURCES