DNA damage in bronchial epithelial and mesothelial cells with and without associated crocidolite asbestos fibers
Jonas Nygren
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Search for more papers by this authorSatu Suhonen
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Search for more papers by this authorCorresponding Author
Hannu Norppa
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Topeliuksenkatu 41 aA, FIN-00250 Helsinki, FinlandSearch for more papers by this authorKaija Linnainmaa
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Search for more papers by this authorJonas Nygren
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Search for more papers by this authorSatu Suhonen
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Search for more papers by this authorCorresponding Author
Hannu Norppa
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Topeliuksenkatu 41 aA, FIN-00250 Helsinki, FinlandSearch for more papers by this authorKaija Linnainmaa
Laboratory of Molecular and Cellular Toxicology, Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland
Search for more papers by this authorAbstract
Mesothelioma is induced almost exclusively by exposure to asbestos fibers. We have investigated whether the induction of DNA damage in human bronchial epithelial BEAS 2B cells and human mesothelial MeT 5A cells by crocidolite asbestos (2 μg/cm2) requires the presence of asbestos fibers in the cells. DNA damage was measured microscopically by the Comet assay, and the presence of fibers in the same cells was assessed using bright-field illumination. After treatment times of 6–72 hr, damage levels were, on the average, two times higher in cells with fibers than in cells without fibers. It was further found that DNA damage decreased with time in BEAS 2B cells both with and without fibers. No decrease in damage with time was seen in MeT 5A cells, suggesting that these mesothelial cells repair the initial damage poorly, lack induction of protective systems, or constantly produce high levels of damaging species. Our results indicate that crocidolite-treated human mesothelial MeT 5A and bronchial epithelial BEAS 2B cells show an elevated level of DNA damage if they contain a fiber. In comparison with epithelial BEAS 2B cells, mesothelial MeT 5A cells have more DNA damage after the crocidolite treatment and the damage is more persistent. Environ. Mol. Mutagen., 2004. © 2004 Wiley-Liss, Inc.
REFERENCES
- Adachi S, Yoshida S, Kawamura K, Takahashi M, Uchida H, Odagiri Y, Takemoto K. 1994. Inductions of oxidative DNA damage and mesothelioma by crocidolite, with special reference to the presence of iron inside and outside of asbestos fiber. Carcinogenesis 15: 753–758.
- Aljandali A, Pollack H, Yeldandi A, Li Y, Weitzman SA, Kamp DW. 2001. Asbestos causes apoptosis in alveolar epithelial cells: role of iron-induced free radicals. J Lab Clin Med 137: 330–339.
- Bocchetta M, Di Resta I, Powers A, Fresco R, Tosolini A, Testa JR, Pass HI, Rizzo P, Carbone M. 2000. Human mesothelial cells are unusually susceptible to simian virus 40-mediated transformation and asbestos cocarcinogenicity. Proc Natl Acad Sci USA 97: 10214–10219.
- Broaddus VC, Yang L, Scavo LM, Ernst JD, Boylan AM. 1997. Crocidolite asbestos induces apoptosis of pleural mesothelial cells: role of reactive oxygen species and poly(ADP-ribosyl) polymerase. Environ Health Perspect 105S: 1147–1152.
- Carbone M, Pass HI, Rizzo P, Marinetti M, Di Muzio M, Mew DJ, Levine AS, Procopio A. 1994. Simian virus 40-like DNA sequences in human pleural mesothelioma. Oncogene 9: 1781–1790.
- Carbone M, Rizzo P, Grimley PM, Procopio A, Mew DJ, Shridhar V, de Bartolomeis A, Esposito V, Giuliano MT, Steinberg SM, Levine AS, Giordano A, Pass HI. 1997. Simian virus-40 large-T antigen binds p53 in human mesotheliomas. Nat Med 3: 908–912.
- Churg A, Keeling B, Gilks B, Porter S, Olive P. 1995. Rat mesothelial and tracheal epithelial cells show equal DNA sensitivity to hydrogen peroxide-induced oxidant injury. Am J Physiol 268: L832–L838.
- Dopp E, Saedler J, Stopper H, Weiss DG, Schiffmann D. 1995. Mitotic disturbances and micronucleus induction in Syrian hamster embryo fibroblast cells caused by asbestos fibers. Environ Health Perspect 103: 268–271.
- Dopp E, Schiffmann D. 1998. Analysis of chromosomal alterations induced by asbestos and ceramic fibers. Toxicol Lett 96–97: 155–162.
- Fenech M. 2000. The in vitro micronucleus technique. Mutat Res 455: 81–95.
- Fung H, Kow YW, Van Houten B, Taatjes DJ, Hatahet Z, Janssen YM, Vacek P, Faux SP, Mossman BT. 1998. Asbestos increases mammalian AP-endonuclease gene expression, protein levels, and enzyme activity in mesothelial cells. Cancer Res 58: 189–194.
- Hart GA, Kathman LM, Hesterberg TW. 1994. In vitro cytotoxicity of asbestos and man-made vitreous fibers: roles of fiber length, diameter and composition. Carcinogenesis 15: 971–977.
- Hesterberg TW, Barrett JC. 1985. Induction by asbestos fibers of anaphase abnormalities: mechanism for aneuploidy induction and possibly carcinogenesis. Carcinogenesis 6: 473–475.
-
Hirvonen A,
Mattson K,
Karjalainen A,
Ollikainen T,
Tammilehto L,
Hovi T,
Vainio H,
Pass HI,
Di Resta I,
Carbone M,
Linnainmaa K.
1999.
Simian virus 40 (SV40)-like DNA sequences not detectable in finnish mesothelioma patients not exposed to SV40-contaminated polio vaccines.
Mol Carcinog
26:
93–99.
10.1002/(SICI)1098-2744(199910)26:2<93::AID-MC4>3.0.CO;2-Z CAS PubMed Web of Science® Google Scholar
- Husgafvel-Pursiainen K, Hackman P, Ridanpää M, Anttila S, Karjalainen A, Partanen T, Taikina-Aho O, Heikkilä L, Vainio H. 1993. K-ras mutations in human adenocarcinoma of the lung: association with smoking and occupational exposure to asbestos. Int J Cancer 53: 250–256.
- Janssen YM, Marsh JP, Absher MP, Gabrielson E, Borm PJ, Driscoll K, Mossman BT. 1994. Oxidant stress responses in human pleural mesothelial cells exposed to asbestos. Am J Respir Crit Care Med 149: 795–802.
- Kamp DW, Graceffa P, Pryor WA, Weitzman SA. 1992. The role of free radicals in asbestos-induced diseases. Free Radic Biol Med 12: 293–315.
- Kamp DW, Israbian VA, Preusen SE, Zhang CX, Weitzman SA. 1995. Asbestos causes DNA strand breaks in cultured pulmonary epithelial cells: role of iron-catalyzed free radicals. Am J Physiol 268: L471–L480.
- Ke Y, Reddel RR, Gerwin BI, Reddel HK, Somers AN, McMenamin MG, LaVeck MA, Stahel RA, Lechner JF, Harris CC. 1989. Establishment of a human in vitro mesothelial cell model system for investigating mechanisms of asbestos-induced mesothelioma. Am J Pathol 134: 979–991.
- Kim HN, Morimoto Y, Tsuda T, Ootsuyama Y, Hirohashi M, Hirano T, Tanaka I, Lim Y, Yun IG, Kasai H. 2001. Changes in DNA 8-hydroxyguanine levels, 8-hydroxyguanine repair activity, and hOGG1 and hMTH1 mRNA expression in human lung alveolar epithelial cells induced by crocidolite asbestos. Carcinogenesis 22: 265–269.
- Kimizuka G, Azuma M, Ishibashi M, Shinozaki K, Hayashi Y. 1993. Co-carcinogenic effect of chrysotile and amosite asbestos with benzo(a)pyrene in the lung of hamsters. Acta Pathol Jpn 43: 149–153.
- Klungland A, Rosewell I, Hollenbach S, Larsen E, Daly G, Epe B, Seeberg E, Lindahl T, Barnes D. 1999. Accumulation of premutagenic DNA lesions in mice defective in removal of oxidative base damage. Proc Natl Acad Sci USA 96: 13300–13305.
- Lieber M, Smith B, Szakal A, Nelson-Rees W, Todaro G. 1976. A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int J Cancer 17: 62–70.
- Levresse V, Renier A, Levy F, Broaddus VC, Jaurand M. 2000. DNA breakage in asbestos-treated normal and transformed (TSV40) rat pleural mesothelial cells. Mutagenesis 15: 239–244.
- Liu W, Ernst JD, Broaddus VC. 2000. Phagocytosis of crocidolite asbestos induces oxidative stress, DNA damage, and apoptosis in mesothelial cells. Am J Respir Cell Mol Biol 23: 371–378.
- McDonald AD, McDonald JC. 1987. Epidemiology of malignant mesothelioma. In: K Antman, J Aisner, editors. Asbestos-related malignancy. Orlando, FL: Grune and Stratton. p 31–55.
- Mossman BT, Eastman A, Landesman JM, Bresnick E. 1983. Effects of crocidolite and chrysotile asbestos on cellular uptake and metabolism of benzo(a)pyrene in hamster tracheal epithelial cells. Environ Health Perspect 51: 331–335.
- Mossman BT, Marsh JP, Shatos MA. 1986. Alteration of superoxide dismutase activity in tracheal epithelial cells by asbestos and inhibition of cytotoxicity by antioxidants. Lab Invest 54: 204–212.
- Mossman BT, Kamp DW, Weitzman SA. 1996. Mechanisms of carcinogenesis and clinical features of asbestos-associated cancers. Cancer Invest 14: 466–480.
- Nyberg P, Klockars M. 1990. Measurement of reactive oxygen metabolites produced by human monocyte-derived macrophages exposed to mineral dusts. Int J Exp Pathol 71: 537–544.
- Ollikainen T, Linnainmaa K, Kinnula VL. 1999. DNA single strand breaks induced by asbestos fibers in human pleural mesothelial cells in vitro. Environ Mol Mutagen 33: 153–160.
- Ollikainen T, Puhakka A, Kahlos K, Linnainmaa K, Kinnula VL. 2000. Modulation of cell and DNA damage by poly(ADP)ribose polymerase in lung cells exposed to H2O2 or asbestos fibres. Mutat Res 470: 77–84.
- Oshimura M, Hesterberg TW, Tsutsui T, Barrett JC. 1984. Correlation of asbestos-induced cytogenetic effects with cell transformation of Syrian hamster embryo cells in culture. Cancer Res 44: 5017–5022.
- Palekar LD, Eyre JF, Most BM, Coffin DL. 1987. Metaphase and anaphase analysis of V79 cells exposed to erionite, UICC chrysotile and UICC crocidolite. Carcinogenesis 8: 553–560.
- Pelin K, Husgafvel-Pursiainen K, Vallas M, Vanhala E, Linnainmaa K. 1992. Cytotoxicity and anaphase aberrations induced by mineral fibres in cultured human mesothelial cells. Toxicol In Vitro 6: 445–450.
- Puhakka A, Ollikainen T, Soini Y, Kahlos K, Saily M, Koistinen P, Pääkkö P, Linnainmaa K, Kinnula VL. 2002. Modulation of DNA single-strand breaks by intracellular glutathione in human lung cells exposed to asbestos fibers. Mutat Res 514: 7–17.
- Reddel RR, Ke Y, Gerwin BI, McMenamin MG, Lechner JF, Su RT, Brash DE, Park JB, Rhim JS, Harris CC. 1988. Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing SV40 early region genes. Cancer Res 48: 1904–1909.
- Singh NP, McCoy MT, Tice RR, Schneider EL. 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175: 184–191.
- Stanton MF, Layard M, Tegeris A, Miller E, May M, Morgan E, Smith A. 1981. Relation of particle dimension to carcinogenicity in amphibole asbestoses and other fibrous minerals. J Natl Cancer Inst 67: 965–975.
- Unfried K, Schurkes C, Abel J. 2002. Distinct spectrum of mutations induced by crocidolite asbestos: clue for 8-hydroxydeoxyguanosine-dependent mutagenesis in vivo. Cancer Res 62: 99–104.
- Vallyathan V, Shi X. 1997. The role of oxygen free radicals in occupational and environmental lung diseases. Environ Health Perspect 105(Suppl 1): 165–177.
- Xu A, Wu LJ, Santella RM, Hei TK. 1999. Role of oxyradicals in mutagenicity and DNA damage induced by crocidolite asbestos in mammalian cells. Cancer Res 59: 5922–5926.