Carcinogenic potential and genomic instability of beryllium sulphate in BALB/c-3T3 cells

  • Nagalakshmi Keshava
  • Gu Zhou
  • Michelle Spruill
  • Mang Ensell
  • Tong-man Ong
Part of the Developments in Molecular and Cellular Biochemistry book series (DMCB, volume 34)

Abstract

Occupational exposure to beryllium (Be) and Be compounds occurs in a wide range of industrial processes. A large number of workers are potentially exposed to this metal during manufacturing and processing, so there is a concern regarding the potential carcinogenic hazard of Be. Studies were performed to determine the carcinogenic potential of beryllium sulfate (BeSO4) in cultured mammalian cells. BALBIc-3T3 cells were treated with varying concentrations of BeSO4for 72 h and the transformation frequency was determined after 4 weeks of culturing. Concentrations from 50-200 jtg BeSO4/m1, caused a concentration-dependent increase (9-41 fold) in transformation frequency. Non-transformed BALB/c-3T3 cells and cells from transformed foci induced by Be5O4were injected into both axillary regions of nude mice. All ten Be-induced transformed cell lines injected into nude mice produced fibrosarcomas within 50 days after cell injection. No tumors were found in nude mice receiving non-transformed BALE/c-3T3 cells 90 days post-injection. Gene amplification was investigated inK-ras,c-myc,c-fos,c jun,c-sis,erb-B2andp53using differential PCR while random amplified polymorphic DNA fingerprinting was employed to detect genomic instability. Gene amplification was found inK-rasandc jun, however no change in gene expression or protein level was observed in any of the genes by Western blotting. Five of the 10 transformed cell lines showed genetic instability using different random primers. In conclusion, these results indicate that BeSO4is capable of inducing morphological cell transformation in mammalian cells and that transformed cells induced by BeSO4are potentially tumorigenic. Also, cell transformation induced by BeSO4may be attributed, in part, to the gene amplification ofK-rasandc-junand some BeSO4-induced transformed cells possess neoplastic potential resulting from genomic instability. (Mol Cell Biochem222:69-76, 2001)

Key words

cell transformation tumorigenicity genomic instability  beryllium sulphate 

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References

  1. 1.
    WHO: Beryllium (Environmental Health Criteria 106). Geneva, 1990Google Scholar
  2. 2.
    IARC Monographs on the Evaluation of the Carcinogenic Risk to Humans: Beryllium, cadmium, mercury, and exposures in the glass manufacturing industry. Lyon 58: 41–117, 1993Google Scholar
  3. 3.
    IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans: Some metals and metallic compounds. Lyon 23: 143–204, 1980Google Scholar
  4. 4.
    Flamm WG: Beryllium: Laboratory evidence. IARC Sci Publ 65: 199–201, 1985PubMedGoogle Scholar
  5. 5.
    Ashby J, Ishidate M Jr, Stoner GD, Morgan MA, Ratpan F, Callander RD: Studies on the genotoxicity of beryllium sulphatein vitroandin vivo.Mutat Res 240: 217–225, 1990PubMedCrossRefGoogle Scholar
  6. 6.
    Dunkel VC, Zeiger E, Brusick D, McCoy E, McGregor D, Mortelmans K, Rosenkranz HS, Simmon VF: Reproducibility of microbial mutagenicity assays: I. Tests withSalmonella typhimuriumandEscherichia coliusing a standardized protocol. Environ Mutagen 6: 1–251, 1984PubMedCrossRefGoogle Scholar
  7. 7.
    Kuroda K, Endo G, Okamoto A, Yo YS, Horiguchi S-1: Genotoxicity of beryllium, gallium and antimony in short-term assays. Mutat Res 264: 163–170, 1991PubMedCrossRefGoogle Scholar
  8. 8.
    Arlauskas A, Baker RS, Bonin AM, Tandon RK, Crisp PT, Ellis J: Mutagenicity of metal ions in bacteria. Environ Res 36: 379–388, 1985PubMedCrossRefGoogle Scholar
  9. 9.
    Larramcndy ML, Popescu NC, DiPaolo JA: Induction by inorganic metal salts of sister chromatid exchanges and chromosome aberration in human and Syrian hamster cell strains. Environ Mutagen 3: 597–606, 1981Google Scholar
  10. 10.
    Miyaki M, Akamatsu N, Ono T, Koyama H: Mutagenicity of metal cations in cultured cells from Chinese hamster. Mutat Res 68: 259–263, 1979PubMedCrossRefGoogle Scholar
  11. 11.
    Dunkel VC, Pienta RJ, Sivak A, Traul KA: Comparative neoplastic transformation responses of Balb/3T3 cells, Syrian hamster embryo cells, and Rauseher murine leukemia virus-infected Fischer 344 rat embryo cells to chemical carcinogens. J Natl Cancer Inst 67: 1303–1315, 1981PubMedGoogle Scholar
  12. 12.
    DiPaolo JA, Casto BC: Quantitative studies ofin vitromorphological transformation of Syrian hamster cells by inorganic metal salts. Cancer Res 39: 1008–1013, 1979PubMedGoogle Scholar
  13. 13.
    Steel VE, Wilkinson BP, Arnold. JT, Kutzman RS: Study of beryllium oxide genotoxicity in cultured respiratory epithelial cells. Inhal Toxicol 1; 95–110, 1989CrossRefGoogle Scholar
  14. 14.
    Finch GL, Hoover MD, Hahn FF, Nikula Ki, Belinsky SA, Haley PJ, Griffith WC: Animal models of beryllium-induced lung disease. Environ Health Perspect 104: 973–979, 1996PubMedGoogle Scholar
  15. 15.
    LeBoeuf RA, Kcrchacrt KA, Aardema MJ, Isfort RJ: The use of short-and medium-term tests for carcinogens and data on genetic effects. In: D.B. McGregor, J.M. Rice, S. Venitt (eds). Carcinogenic Hazard Evaluation. IARC Sci Publ 146: 409–425, 1999Google Scholar
  16. 16.
    Stutman O: In: J. Fogh, B. Giovanella (eds). The Nude Mouse in Experimental and Clinical Research. Academic Press, New York, 1978, pp 411–435Google Scholar
  17. 17.
    Dunkel VC, Rogers C, Swierenga SHH, Brillinger RL, Gilman JPW, Nestmann ER: Reconunended protocols based on a survey of current practice in genotoxicity testing laboratories: III. Cell transformation in C3H11 OT 112 mouse embryo cell, BALB/c 3T3 mouse fibroblast and Syrian hamster embryo cell cultures. Mutat Res 246: 285–300, 1991PubMedCrossRefGoogle Scholar
  18. 18.
    Sambrook J, Fritsch EF, Maniatis T: In: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1980Google Scholar
  19. 19.
    Niman HL, Thompson AMH, Yu A, Markham M, Willems JJ, Herwig KR, Habib NA, Wood CB: Antipeptide antibodies to detect oncogenerelated proteins in urine. Proc Natl Aced Sci USA 82: 7924–7928, 1985CrossRefGoogle Scholar
  20. 20.
    Simon VF: Invitromulagenicity assays of chemical carcinogens and related compounds withSalmonella typhimurium.J Natl Cancer lnst 62: 911–918, 1979Google Scholar
  21. 21.
    Tso WW, Fung WP: Mutagenicity of metallic cations. Toxicol Lett 8: 195–200,1981PubMedCrossRefGoogle Scholar
  22. 22.
    Tu AS, Murray TA, Hatch KM, Sivak A, Milman HA: Invitrotransformation of BALBIc-3T3 cells by chlorinated ethanes and ethylenes. Cancer Lett 28: 85–92, 1985PubMedCrossRefGoogle Scholar
  23. 23.
    Fitzgerald DJ, Piccoli C, Yamasaki H: Detection of non-gcnotoxic carcinogens in the BALB/c 3T3 cell transformation/mutation assay system. Mutagenesis 4: 286–29 I, 1989CrossRefGoogle Scholar
  24. 24.
    Colacci A, Vaccari M, Peroceo P, Via CD, Silingardi P, Manzin E, Horn W, Grilli S: Enhancement of BALB/c 3T3 cells transformation by 1,2dibromoethanc promoting effect. Carcinogenesis 17: 225–231, 1996PubMedCrossRefGoogle Scholar
  25. 25.
    MacDonald F, Ford Ci: In: Molecular Biology of Cancer. Bios Scientific Publisher, Oxford, 1997Google Scholar
  26. 26.
    Cooper G, Okenquist S, Silverman L: Transforming activity of DNA of chemically transformed and normal cells. Nature 284: 418 421, 1980Google Scholar
  27. 27.
    Berwald Y, Sachs L:In vitrotransformation with chemical carcinogens. Nature 200: 1182–1184, 1963PubMedCrossRefGoogle Scholar
  28. 28.
    Isford RJ, LeBoeuf RA: The Syrian hamster embryo (SHE) cell transformation system: A biologically relevant in vitro model — with carcinogen predicting capabilities — of in vivo multistage neoplastic transformation. Crit Rev Oncol 6: 251–260, 1995CrossRefGoogle Scholar
  29. 29.
    Belinsky SA, Swafford DS, Finch GL, Mitchell CE, Kelly G, Hahn FF, Anderson MW, Nikula KJ: Alterations in the K-ras and p53 genes in rat lung tumors. Environ Health Perspect 105: 901–906, 1997PubMedGoogle Scholar
  30. 30.
    Nickell-Brady C, Hahn FF, Finch GL, Belinsky SA: Analysis of K-ras, p53 and c-raf-1 mutations in beryllium-induced rat lung tumors. Carcinogenesis 15: 257–262, 1994PubMedCrossRefGoogle Scholar
  31. 31.
    Finch GL, Hoover MD, Hahn FF, Nikula KJ, Belinsky SA, Haley PJ, Griffith WC: Animal models of beryllium-induced lung disease. Environ Health Perspect 104: 973–979, 1996PubMedGoogle Scholar
  32. 32.
    Gao H-G, Brick I, Ong S, Miller M, Whong W-Z, Ong T: Selective hyperexpression ofc junoncoprotein by glass fiber-and silica-transformed BALBIc3T3 cells. Cancer Lett 112: 65–69, 1997PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2001

Authors and Affiliations

  • Nagalakshmi Keshava
    • 1
  • Gu Zhou
    • 1
  • Michelle Spruill
    • 1
  • Mang Ensell
    • 1
  • Tong-man Ong
    • 1
  1. 1.Health Effects Laboratory DivisionNational Institute for Occupational Safety and Health, Centers for Disease Control and PreventionMorgantownUSA

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