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Telomerase-mediated lifespan extension of human bronchial cells does not affect hexavalent chromium-induced cytotoxicity or genotoxicity

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Abstract

Hexavalent chromium (Cr(VI)) is a metal of increasing public health concern, as exposure to it is widespread and it is a well-established cause of human bronchial carcinomas and fibrosarcomas. The water-insoluble Cr(VI) salts are potent carcinogens compared to the water soluble salts; yet the genotoxic mechanisms of both may be mediated by soluble Cr(VI) ions. Currently, these mechanisms are poorly understood. Emerging evidence suggests that initial cell culture models used to study the general toxicity of Cr(VI) may be suboptimal for investigating mechanisms specific to human bronchial cells. Accordingly, we have developed a new model system of human bronchial cells by introducing hTERT, the catalytic subunit of human telomerase, into primary human bronchial fibroblasts (PHBF). We have isolated a stable, clonally derived cell line, WHTBF-6, that demonstrate reconstitution of telomerase activity and maintenance of telomere lengths with increasing culture age. WHTBF-6 has been characterized as having an extended in vitro lifespan, a normal growth rate, a normal diploid karyotype that is maintained over time, and exhibits serum-dependent contact-inhibited anchorage-dependent growth. Moreover, we find that both particulate and soluble hexavalent chromium induce a pattern and degree of cytotoxicity and clastogenicity in WHTBF-6 that is similar to the parental PHBF cells. Because telomerase does not compromise growth or the response to Cr(VI), our results indicate that this is an excellent system for studying the mechanisms of Cr(VI) and potentially other carcinogens implicated in the development of lung cancer.

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References

  1. Agency for Toxic Substances and Disease Research: Top 20 hazardous substances: ATSDR/EPA priority list for 1997. U.S. Department of Health and Human Services Public Health Service/U.S. Environmental Protection Agency

  2. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans: Chromium, Nickel and Welding, Vol. 49. International Agency for Research on Cancer, Lyons, France, 1990

    Google Scholar 

  3. National Occupational Exposure Survey, National Institute for Occupational Safety and Health, Cincinnati, Ohio, 1988

  4. Johnson BL, DeRosa CT: The toxicological hazard of superfund hazardous waste sites. Rev Environ Health 12: 235–251, 1997

    Google Scholar 

  5. Damage and Threats caused by Hazardous Materials Sits, EPA/430-9-80/004, U.S. Environmental Protection Agency, Washington DC, 1980

  6. Léonard A, Lauwerys RR: Carcinogenicity and mutagenicity of chromium. Mutat Res 76: 239–277, 1980

    Google Scholar 

  7. Levy LS, Vanitt S: Carcinogenicity and mutagenicity of chromium compounds: The association between bronchial metaplasia and neoplasia. Carcinogenesis 7: 831–835, 1986

    Google Scholar 

  8. Patierno SR, Banh D, Landolph JR: Transformation of C3H/10T1/2 mouse embryo cells by insoluble lead chromate but not soluble calcium chromate: Relationship to mutagenesis and internalization of lead chromate particles. Cancer Res 47: 3815–3823, 1988

    Google Scholar 

  9. Ishikawa Y, Nakagawa K, Satoh Y, Kitagawa T, Sugano H, Hirano T, Tsuchiya E: Characteristics of chromate workers' cancers, chromium lung deposition and precancerous bronchial lesions: An autopsy study. Br J Cancer 70: 160–166, 1994

    Google Scholar 

  10. Ishikawa Y, Nakagawa K, Satoh Y, Kitagawa T, Sugano H, Hirano T, Tsuchiya E: ‘Hot spots’ of chromium accumulation at bifurcations of chromate workers' bronchi. Cancer Res 2342–2346, 1994

  11. DeFlora S, Bagnasco M, Serra D, Zanacchi P: Genotoxicity of chromium compounds. A review. Mutat Res 238: 99–172, 1990

    Google Scholar 

  12. Wise JP, Leonard JC, Patierno SR: Clastogenesis by particulate PbCrO4 in hamster and human cells. Mutat Res 278: 69–79, 1992

    Google Scholar 

  13. Wise JP, Orenstein JM, Patierno SR: Inhibition of lead chromate clastogenesis by ascorbate: Relationship to particle dissolution and uptake. Carcinogenesis 14: 429–434, 1993

    Google Scholar 

  14. Wise JP Sr, Stearns DM, Wetterhahn KE, Patierno SR: Cell-enhanced dissolution of carcinogenic lead chromate particles: The role of individual dissolution products in clastogenesis. Carcinogenesis 15: 2249–2254, 1994

    Google Scholar 

  15. Lechner JF, LaVeck MA: A serum-free method for culturing normal human bronchial epithelial cells at clonal density. J Tissue Cult Meth 9: 43–48, 1985

    Google Scholar 

  16. Wise JP Sr, Wise SS, Little JE: The cytotoxicity and genotoxicity of particulate and soluble hexavalent chromium in human lung cells. Mutat Res 517: 221–229, 2002

    Google Scholar 

  17. Singh J, Pritchard DE, Carlisle DL, McLean JA, Montaser A, Orenstein JM, Patierno SR: Internalization of carcinogenic lead chromate particles by cultured normal human lung epithelial cells. Formation of intracellular lead-inclusion bodies and induction of apoptosis. Toxicol Appl Pharm 161: 240–248, 1999

    Google Scholar 

  18. Fornace AJ Jr, Seres DS, Lechner JF, Harris CC: DNA-protein cross-linking by chromium salts. Chem Biol Interact. 36: 345–354, 1981

    Google Scholar 

  19. Xu J, Bubley GJ, Detrick B, Blankenship LJ, Patierno SR: Chromium(VI) treatment of normal human lung cells results in guanine-specific DNA polymerase arrest, DNA-DNA cross-links and S-phase blockade of cell cycle. Carcinogenesis 17: 1511–1517, 1996

    Google Scholar 

  20. Carlisle DL, Pritchard DE, Singh J, Owens BM, Blankenship LJ, Orenstein JM, Patierno SR: Apoptosis and p53 induction in human lung fibroblasts exposed to chromium(VI): Effect of ascorbate and tocopherol. Toxicol Sci 55: 60–68, 2000

    Google Scholar 

  21. Carlisle DL, Pritchard DE, Singh J, Patierno SR: Chromium(VI) induces p53-dependent apoptosis in diploid human lung and mouse dermal fibroblasts. Mol Carcinogen 28: 111–118, 2000

    Google Scholar 

  22. Reddel RR, Ke Y, Gerwin BI, McMenamin MG, Lechner JR, Su RT, Brash DE, Park JB, Rhim JS, Harris CC: 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, 1988

    Google Scholar 

  23. Willey JC, Broussoud A, Sleemi A, Bennett WP, Cerutti P, Harris CC: Immortalization of normal human bronchial epithelial cells by human papillomaviruses 16 or 18. Cancer Res 51: 5370–5377, 1991

    Google Scholar 

  24. Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu C-P, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE: Extension of life-span by introduction of telomerase into normal human cells. Science 207: 349–352, 1998

    Google Scholar 

  25. MacKenzie KL, Franco S, May C, Sadelain M, Moore MA: Mass cultured human fibroblasts overexpressing hTERT encounter, a growth crisis following an extended period of proliferation. Exp Cell Res 259: 336–350, 2000

    Google Scholar 

  26. Ren JG, Xia HL, Tian YM, Just T, Cai GP, Dai YR: Expression of telomerase inhibits hydroxyl radical-induced apoptosis in normal telomerase negative human lung fibroblasts. FEBS Lett 488: 133–138, 2001

    Google Scholar 

  27. Harley CB, Futcher AB, Greider CW: Telomeres shorten during ageing of human fibroblasts. Nature 345: 458–460, 1990

    Google Scholar 

  28. Hastie N, Dempster M, Dunlop MG, Thompson AM, Green DK, Allshire RC: Telomere reduction in human colorectal carcinoma and with ageing. Nature 346: 866–868, 1990

    Google Scholar 

  29. Weinrich SL, Pruzan R, Ma L, Ouellette M, Tesmer VM, Holt SE, Bodnar AG, Lichtsteiner S, Kim NW, Trager JB, Taylor RD, Carlos R, Andrews WH, Wright WE, Shay JW, Harley CB, Morin GB: Reconstitution of human telomerase with the template RNA component hTR and the catalytic protein subunit hTRT. Nature Genet 17: 498–502, 1997

    Google Scholar 

  30. Nakamura TM, Morin GB, Chapman KB, Weinrich SL, Andrews WH, Lingner J, Harley CB, Cech TR: Telomerase catalytic subunit homologs from fission yeast and human. Science 277: 955–959, 1997

    Google Scholar 

  31. Jiang X, Jimenez G, Chang E, Frolkis M, Kusler B, Sage M, Beeche M, Bodnar AG, Wahl GM, Tlsty TD, Chiu C: Telomerase expression in human somatic cells does not induce changes associated with a transformed phenotype. Nature Genet 21: 111–114, 1999

    Google Scholar 

  32. Morales CP, Holt SE, Ouellette M, Kaur KJ, Yan Y, Wilson KS, White A, Wright WE, Shay JW: Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nature Genet 21: 115–118, 1999

    Google Scholar 

  33. Vaziri H, Squire JA, Pandita TK, Bradley G, Kuba RM, Zhang H, Gulyas S, Hill RP, Nolan GP, Benchimol S: Analysis of genomic integrity and p53-dependent G1 checkpoint in telomerase-induced extended-life-span human fibroblasts. Mol Cell Biol 19: 2373–2379, 1999

    Google Scholar 

  34. Savre-Train I, Gollahon LS, Holt SE: Clonal heterogeneity in telomerase activity and telomere length in tumor-derived cell lines. Proc Soc Exp Biol Med 223: 379–388, 2000

    Google Scholar 

  35. Holt SE, Norton JC, Wright WE, Shay JW: Comparison of the telomeric repeat amplification protocol (TRAP) to the new TRAP-eze telomerase detection kit. Meth Cell Sci 18: 237–248, 1996

    Google Scholar 

  36. Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medranos EE, Linskens M, Rubelj I, Pereira-Smith O, Peacocke M, Campisi J: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 92: 9363–9367, 1995

    Google Scholar 

  37. Beattie TL, Zhou W, Robinson M, Harrington L: Reconstitution of human telomerase activity in vitro. Curr Biol 8: 117–180, 1998

    Google Scholar 

  38. Dubrovskaya VA, Wetterhahn KE: Effects of CR(VI) on the expression of the oxidative stress genes in human lung cells. Carcinogenesis 19: 1401–1407, 1998

    Google Scholar 

  39. Bidulescu A, Wise SS, May A, Tang ER, Bryant BH, Schumm KR, Wise JP Sr: Hexavalent chromium toxicity in human lung cells, Chinese hamster ovary cells and A549 cells: Current cell culture models are inadequate. Toxicol Sci 66: 32, 2002

    Google Scholar 

  40. Ouellette MM, Liao M, Herbert B-S, Johnson M, Holt SE, Liss HS, Shay JW, Wright WE: Subsenescent telomere lengths in fibroblasts immortalized by limiting amounts of telomerase. J Biol Chem 275: 10072–10076, 2000

    Google Scholar 

  41. Pritchard DE, Ceryak S, Ha L, Fornsaglio JL, Hartman SK, O'Brien TJ, Patierno SR: Mechanism of apoptosis and determination of cellular fate in chromium(VI)-exposed populations of telomerase immortalized fibroblasts. Cell Growth Diff 12: 347–363, 2001

    Google Scholar 

  42. Yunis JJ: The chromosomal basis for human neoplasia. Science 221: 227–236, 1983

    Google Scholar 

  43. Cavanee WK, Dryja TP, Phillips RA, Benedict WF, Godbout R, Gallie BL, Murphee AL, Strong LC, White RL: Expression of recessive alleles by chromosomal mechanisms in retinoblastoma. Nature 305: 779–784, 1983

    Google Scholar 

  44. Adams J, Gerondakis S, Webb E, Corcoran LM, Cory S: Cellular myc oncogene is altered by chromosome translocation to an immunoglobulin locus in murine plasmacytomas and is rearranged similarly in human Burkitt lymphomas. Proc Natl Acad Sci USA 80: 1982–1986, 1983

    Google Scholar 

  45. Davis RL, Konopka JB, Witte ON: Activation of the c-abl oncogene by viral transduction or chromosomal translocation generates altered c-abl proteins with similar in vitro kinase properties. Mol Cell Biol 5 204–213, 1985

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Environmental and Genetic Toxicology, Department of Epidemiology and Public Health, Division of Environmental Health Sciences, Yale University School of Medicine, New Haven, CT, USA

    Sandra S. Wise, Jennifer E. Little, Peter G. Antonucci & Bronwyn H. Bryant

  2. Department of Pathology, Medical College of Virginia at Virginia Commonwealth University, Richmond, VA, USA

    Lynne W. Elmore & Shawn E. Holt

  3. Center for Integrated and Applied Environmental Toxicology, University of Southern Maine, PO Box 9300, Portland, Maine, 04104-9300, USA

    John Pierce Wise Sr

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  1. Sandra S. Wise
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  2. Lynne W. Elmore
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  7. John Pierce Wise Sr
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Wise, S.S., Elmore, L.W., Holt, S.E. et al. Telomerase-mediated lifespan extension of human bronchial cells does not affect hexavalent chromium-induced cytotoxicity or genotoxicity. Mol Cell Biochem 255, 103–112 (2004). https://doi.org/10.1023/B:MCBI.0000007266.82705.d9

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  • DOI: https://doi.org/10.1023/B:MCBI.0000007266.82705.d9

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