Molecular and Cellular Biochemistry

, Volume 255, Issue 1–2, pp 247–256 | Cite as

Effect of the militarily-relevant heavy metals, depleted uranium and heavy metal tungsten-alloy on gene expression in human liver carcinoma cells (HepG2)

  • Alexandra C. Miller
  • Kia Brooks
  • Jan Smith
  • Natalie Page

Abstract

Depleted uranium (DU) and heavy-metal tungsten alloys (HMTAs) are dense heavy-metals used primarily in military applications. Chemically similar to natural uranium, but depleted of the higher activity 235U and 234U isotopes, DU is a low specific activity, high-density heavy metal. In contrast, the non-radioactive HMTAs are composed of a mixture of tungsten (91–93%), nickel (3–5%), and cobalt (2–4%) particles. The use of DU and HMTAs in military munitions could result in their internalization in humans. Limited data exist however, regarding the long-term health effects of internalized DU and HMTAs in humans. Both DU and HMTAs possess a tumorigenic transforming potential and are genotoxic and mutagenic in vitro. Using insoluble DU-UO2 and a reconstituted mixture of tungsten, nickel, cobalt (rWNiCo), we tested their ability to induce stress genes in thirteen different recombinant cell lines generated from human liver carcinoma cells (HepG2). The commercially available CAT-Tox (L) cellular assay consists of a panel of cell lines stably transfected with reporter genes consisting of a coding sequence for chloramphenicol acetyl transferase (CAT) under transcriptional control by mammalian stress gene regulatory sequences. DU, (5–50 μg/ml) produced a complex profile of activity demonstrating significant dose-dependent induction of the hMTIIA FOS, p53RE, Gadd153, Gadd45, NFκBRE, CRE, HSP70, RARE, and GRP78 promoters. The rWNiCo mixture (5–50 μg/ml) showed dose-related induction of the GSTYA, hMTIIA, p53RE, FOS, NFκBRE, HSP70, and CRE promoters. An examination of the pure metals, tungsten (W), nickel (Ni), and cobalt (Co), comprising the rWNiCo mixture, demonstrated that each metal exhibited a similar pattern of gene induction, but at a significantly decreased magnitude than that of the rWNiCo mixture. These data showed a synergistic activation of gene expression by the metals in the rWNiCo mixture. Our data show for the first time that DU and rWNiCo can activate gene expression through several signal transduction pathways that may be involved in the toxicity and tumorigenicity of both DU and HMTAs.

depleted uranium heavy-metals tungsten alloy metals HEPG2 cells gene expression 

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References

  1. 1.
    Miller AC, Blakely WF, Livengood D, Whittaker T, Xu J, Ejnik JW, Hamilton MM, Parlette E, St. John T, Gerstenberg HM, Hsu H: Transformation of human osteoblast cells to the tumorigenic phenotype by depleted uranium-uranyl chlroide. Environ Health Perspect 106: 465–471, 1998Google Scholar
  2. 2.
    Miller AC, Fuciarelli AF, Jackson WE, Ejnik EJ, Emond C, Strocko S, Hogan J, Page N, Pellmar T: Urinary and serum muatagenicity studies with rats implanted with depleted uranium or tantalum pellets. Mutagenesis 13: 101–106, 1998Google Scholar
  3. 3.
    Andrew SP, Caligiuri RD, Eiselstein LE: A review of penetration mechanisms and dynamic properties of tungsten and depleted uranium penetrators. In: A. Crowson, E.S. Chen (eds). Tungsten and Tungsten Alloys: Recent Advances. Plenum Press, New York, 1991Google Scholar
  4. 4.
    Miller AC, Whittaker T, Hogan J, McBride S, Benson K: Oncogenes as biomarkers for low dose radiation-induced health effects. Can Detect Prev 20: 235–236, 1996Google Scholar
  5. 5.
    Pellmar TC, Fuciarelli AF, Ejnik JW, Hamilton M, Hogan J, Strocko S, Emond C, Mottaz HM, Landauer MR: Distribution of uranium in rats implanted with depleted uranium pellets. Toxicol Sci 49: 29–39, 1999Google Scholar
  6. 6.
    Pellmar TC, Kaiser DO, Emond C, Hogan JB: Electrophysiological changes in hippocampal slices isolated from rats embedded with depleted uranium fragments. Neurotoxicol 20: 785–792, 1999Google Scholar
  7. 7.
    Miller AC, Xu J, Mog S, McKinney L, Page N: Neoplastic transformation of human osteoblast cells to the tumorigenic phenotype by heavy metal-tungsten alloy particles: Induction of genotoxic effects. Carcinogenesis 22: 115–125, 2001Google Scholar
  8. 8.
    Miller AC, Brooks K, Stewart M, Shi L, McClain D, Page N: Genomic instability in human osteoblast cells after exposure to depleted uranium: Delayed lethality and micronucleus formation. J Environ Radioact 64: 247–259, 2003Google Scholar
  9. 9.
    Cugell DW, Morgan WK, Perkins DSG, Rubin A: The respiratory effects of cobalt. Arch Intern Med 150: 177–183, 1990Google Scholar
  10. 10.
    Lauwerys R, Lison D: Health risks associated with cobalt exposure — an overview. Sci Tot Environ 150: 1–6, 1994Google Scholar
  11. 11.
    Lison D, Lauwreys R: The interaction of cobalt metal with different carbides and other mineral particles on mouse peritoneal macrophages. Toxic In Vitro 9: 341–347, 1995Google Scholar
  12. 12.
    Lison D, Carbonelle P, Mollo L, Lauwerys R, Fubini B: Physiochemical mechanism of the interaction between cobalt metal and carbide particles to generate toxic activated oxygen species. Chem Res Toxicol 8: 600–606, 1995Google Scholar
  13. 13.
    Vincent R, Goegan P, Johnson G, Brook JR: Kumarathasan P, Bouthillier L, Burnett R: Regulation of promoter-CAT stress genes in HepG2 cells by suspensions of particles from ambient air. Fundament Appl Toxicol 39: 18–32, 1997Google Scholar
  14. 14.
    Tully DB, Collins BJ, Overstreet JD, Smith CS, Dinse GE, Mumtaz MM, Chapin RE: Effects of arsenic, cadmium, chromium, and lead on 13 different promoters in recombinant HepG2 cells. Toxicol Appl Pharmacol 168: 79–90, 2000Google Scholar
  15. 15.
    Tchounwou PB, Wison BA, Ishaque AB, Schneider J: Atrazine potentiation of arsenic-trioxide-induced cytotoxicity and gene expression in human liver carcinoma cells (HepG2). Mol Cell Biochem 222: 49–59, 2001Google Scholar
  16. 16.
    Miller AC, Xu J, Stewart M, Emond C, Hodge S, Matthews CR, Kalinich J, McClain D: Potential health effects of the heavy metals, depleted uranium and tungsten, used in armor-piercing munitions: Comparison of neoplastic transformation, mutagenicity, genomic instability, and oncogenesis. Metal Ions in Biol Med 6: 209–211, 2000.Google Scholar
  17. 17.
    Miller AC, Stewart M, Brooks K, Shi L, Page N: Depleted-uranium catalyzed oxidative DNA damage: Absence of significant alpha particle decay. J Inorg Biochem 91: 246–252, 2002Google Scholar
  18. 18.
    Miller AC, Xu J, Stewart J, Brooks K, Hodge S, Shi L, Anderson BL, Page N, McClain D: Observation of radiation-specific damage in human cells exposed to depleted uranium: Dicentric frequency and neoplastic transformation as endpoints. Radiat Protect Dosimetry 99: 275–278, 2002Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Alexandra C. Miller
    • 1
  • Kia Brooks
    • 1
  • Jan Smith
    • 2
  • Natalie Page
    • 2
  1. 1.Applied Cellular Radiobiology Department, Armed Forces Radiobiology Research InstituteBethesdaUSA
  2. 2.Molecular Oncology Branch, Division of Cancer Treatment, National Can er Institute, National Institute of HealthBethesdaUSA

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