Carcinogen Metabolism in Immortalised Human Cells Grown as Hybrid Cells in Culture

  • Stuart Brown
  • Helen Ross
Part of the NATO ASI Series book series (NSSA, volume 218)


Most cells from normal tissues have two characteristic properties. Their cell division is regulated in a particular way and they produce substances characteristic only of their tissue of origin. It was soon realised, however, that as cells from normal tissues were studied extensively, a limit to their long term cultivation was found. Normal cells died after a finite number of divisions. In contrast, tumour cells grew indefinitely in culture and usually did not express differentiated functions. In order to distinguish between these types of cells with finite or infinite lifespan in culture, Hayflick and Moorhead (1) used the term cell strain to denote normal cells with a finite lifespan and reserved the term cell line for cells which were established in culture and would divide indefinitely. They also noted that the property of infinite cell growth was usually associated with a change in the diploid nature of the cells and that a heteroploid karyotype was common in permanent cell lines.


Hybrid Cell Epoxide Hydrolase Aryl Hydrocarbon Hydroxylase Permanent Cell Line Aryl Hydrocarbon Hydroxylase Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    L. Hayflick and P.S. Moorhead, The serial cultivation of human diploid cell strains, Exp. Cell Res. 25: 585–621 (1961).CrossRefGoogle Scholar
  2. 2.
    O.M. Pereira-Smith and J.R. Smith, Evidence for the recessive nature of cellular immortality, Science 221: 964–966 (1983).PubMedCrossRefGoogle Scholar
  3. 3.
    H.E. Ruley, Adenovirus early region 1A enables viral and cellular transforming genes to transform primary cells in culture, Nature 304: 602–606 (1983).PubMedCrossRefGoogle Scholar
  4. 4.
    P. van den Elsen, A. Houweling and A.J. van der Eb, Expression of region E1B of human adenovirus in the absence of region EIA is not sufficient for complete transformation, Virology 128: 377–390 (1983).PubMedCrossRefGoogle Scholar
  5. 5.
    B. Zerler, B. Moran, K. Maruyama, J. Moomaw, T. Grodzicker and H.E. Ruley, Adenovirus EIA coding sequences that enable ras and pmt oncogenes to transform cultured primary cells, Mol. Cell Biol. 6: 887–899 (1986).PubMedGoogle Scholar
  6. 6.
    R. Cone, T. Grodzicker and M. Jaramillo, A retrovirus expressing the 12S adenoviral EIA gene products can immortalise epithelial cells from a broad range of rat tissues, Mol. Cell. Biol. 8: 1036–1044 (1988).PubMedGoogle Scholar
  7. 7.
    P. Whyte, K.J. Buchkovich, J.M. Horowitz, S.H. Friend, M. Raybuck, R.A. Weinberg and E. Harlow, Association between an oncogene and an anti-oncogene: the adenovirus EIA proteins bind to the retinoblastoma gene product, Nature 334: 124–129 (1988).PubMedCrossRefGoogle Scholar
  8. 8.
    S-P. Yee and P.E. Branton, Detection of cellular proteins associated with human adenovirus type 5 early region EIA polypeptides, Virology 147: 142–153 (1985).PubMedCrossRefGoogle Scholar
  9. 9.
    E. Harlow, P. Whyte, B.R. Franza and C. Schley, Association of adenovirus early-region 1A proteins with cellular polypeptides, Mol. Cell Biol. 6: 1579–1589 (1986).PubMedGoogle Scholar
  10. 10.
    N. Dyson, P.M. Howley, K. Munger and E. Harlow, The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product, Science 243: 934–937 (1989).PubMedCrossRefGoogle Scholar
  11. 11.
    P. Clertant and I. Seif, A common function for polyoma virus large T and papillomavirus E1 proteins, Nature 311: 276–279 (1984).PubMedCrossRefGoogle Scholar
  12. 12.
    M.E. Ewen, J.W. Ludlow, E. Marsillo, J.A. DeCaprio, R.C. Millikan, S.H. Cheng, E. Paucha and D.M. Livingston, An N-terminal transformation-governing sequence of SVAO large T antigen contributes to the binding of both p110Rb and a second cellular protein, p120, Cell 58: 257–267 (1989).PubMedCrossRefGoogle Scholar
  13. 13.
    M.D. Waterfield, G.T. Scrace, N. Whittle, P. Stroobant, A. Johnsson, A. Wasteson, B. Westermark, C-H. Heldin, J.S. Huang and T.F. Deuel, Platelet derived growth factor is structurally related to the putative transforming protein p28SIS of simian sarcoma virus, Nature 304: 35–39 (1983).PubMedCrossRefGoogle Scholar
  14. 14.
    R.F. Doolittle, M.W. Hunkapiller, L.E. Hood, S.G. Devare, K.C. Robbins, S.A. Aaronson and H.N. Antoniades, Simian Sarcoma Virus onc. gene, v-sis, is derived from the gene (or genes) encoding a platelet derived growth factor, Science 221: 275–277 (1983).PubMedCrossRefGoogle Scholar
  15. 15.
    J. Downward, Y. Yarden, E. Mayes, G. Scrace, N. Totty, P. Stockwell, A. Ullrich, J. Schlessinger and M.D. Waterfield, Close similarity of epidermal growth factor receptor and v-erb B oncogene protein sequences, Nature 307: 521–527 (1984).PubMedCrossRefGoogle Scholar
  16. 16.
    M.S. Collett and R.L. Erikson, Protein kinase activity associated with the avian sarcoma virus src gene product, Proc. Natl. Acad. Sci. U.S.A. 75: 2021–2024 (1978).PubMedCrossRefGoogle Scholar
  17. 17.
    A. Ullrich, L. Coussens, J.S. Hayflick, T.J. Dull, A. Gray, A.W. Tarn, J. Lee, Y. Yarden, T.A. Liebermann, J. Schlessinger, J. Downward, E.L.V. Mayes, N. Whittle, M.D. Waterfield and P.H. Seeburg, Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells, Nature 309: 418–425 (1984).PubMedCrossRefGoogle Scholar
  18. 18.
    J.M. Bishop, Tricks with tyrosine kinases, Nature 319: 722–723 (1986).PubMedCrossRefGoogle Scholar
  19. 19.
    A. Balamin, Transforming ras oncogenes and multistage carcinogenesis, Brit. J. Cancer 51: 1–7 (1985).CrossRefGoogle Scholar
  20. 20.
    R.A. Weinberg, The action of oncogenes in the cytoplasm and nucleus, Science 230: 770–776 (1985).PubMedCrossRefGoogle Scholar
  21. 21.
    K. Kelly, B.H. Cochran, C.D. Stiles and P. Leder, Cell-specific regulation of the c-myc gene by lymphocyte mitogens and platelet derived growth factor, Cell 35: 603–610 (1983).PubMedCrossRefGoogle Scholar
  22. 22.
    J.M. Blanchard, M. Piechaczyk, C. Dani, J.C. Chambard, A. Franchi, J. Pouyssegur and P. Jeanteur, C-myc gene is transcribed at high rate in Go arrested fibroblasts and is post transcriptionally regulated in response to growth factors, Nature 317: 443–445 (1985).PubMedCrossRefGoogle Scholar
  23. 23.
    K.L. Fink, E.D. Wieben, G.E. Woloschak and T.C. Speisberg, Rapid regulation of c-myc protooncogene expression by progesterone in the avian oviduct, Proc. Natl. Acad. Sci. U.S.A. 85: 1796–1800 (1988).PubMedCrossRefGoogle Scholar
  24. 24.
    D.L. Bentley and M. Groudine, A block to elongation is largely responsible for decreased transcription of c-myc in differentiated HL60 cells, Nature 321: 702–706 (1986).PubMedCrossRefGoogle Scholar
  25. 25.
    P. Nath, R. Getzenberg, D. Beebe, L. Pallansch and P. Zelenka, c-myc mRNA is elevated as differentiating lens cells withdraw from the cell cycle, Exp. Cell Res. 169: 215–222 (1987).PubMedCrossRefGoogle Scholar
  26. 26.
    N Sawada, Hepatocytes from old rats retain responsiveness of c-myc expression to EGF in primary culture but do not enter S phase, Exp. Cell Res. 181: 584–588 (1989).PubMedCrossRefGoogle Scholar
  27. 27.
    R.F. Newbold, R.W. Overell and J.R. Connell, Induction of immortality is an early event in malignant transformation of mammalian cells by carcinogens, Nature 299: 633–635 (1982).PubMedCrossRefGoogle Scholar
  28. 28.
    R.T. Su and Y-C. Chang, Transformation of human epidermal cells by transfection with plasmid containing Simian Virus 40 DNA linked to a neomycin gene in a defined medium, Exp. Cell Res. 180: 117–133 (1989).PubMedCrossRefGoogle Scholar
  29. 29.
    G. Kohler and C. Milstein, Continuous cultures of fused cells secreting antibody of predefined specificity, Nature 256: 495–497 (1975).PubMedCrossRefGoogle Scholar
  30. 30.
    L. Olsson and H.S. Kaplan, Human-human monoclonal antibody producing hybridomas: technical aspects, in Methods in Enzymology, vol. 92 pp. 3–16, eds. J.J. Langone and H. van Vunakis, Acad. Press, N.Y. (1983).Google Scholar
  31. 31.
    D.H. Bissell and P.A. Guzelian, Microsomal functions and phenotypic change in adult rat hepatocytes in primary monolayer cultures, in Gene expression and Carcinogenesis in Cultured Liver, ed. L.E. Gerschenson and E.B. Thompson, pp. 119–136, Acad. Press, N.Y. (1975).Google Scholar
  32. 32.
    O. Pelkonen and D.W. Nebert, Metabolism of Polycyclic Aromatic Hydrocarbons: Etiologic role in carcinogenesis, Pharmacol. Revs. 34: 189–251 (1982).Google Scholar
  33. 33.
    B. Paigen, H.L. Gurtoo, J. Minowada, E. Ward, L. Houten, K. Paigen, A. Reilly and R. Vincent, Genetics of aryl hydrocarbon hydroxylase in the human population and its relationship to lung cancer, in Polycyclic Hydrocarbons and Cancer, ed. H.V. Gelboin and P.O.P. Ts’O, pp. 391–406, Acad. Press, N.Y. (1978).Google Scholar
  34. 34.
    P. Wang, J. Meijer and F.P. Guengerich, Purification of human liver cytosolic epoxide hydrolase and comparison to the microsomal enzyme, Biochemistry 21: 5769–5776 (1982).PubMedCrossRefGoogle Scholar
  35. 35.
    C.B. Kasper and D. Henton, Enzymatic basis of detoxification, vol. 2, pp. 3–36, ed. W.B. Jakoby, Acad. Press, N.Y. (1980).Google Scholar
  36. 36.
    B. Felluga, A. Claude and E. Mrena, Electron microscope observations on virus particles associated with a transplantable renal adenocarcinoma in BALB/cf/cd mice, J. Natl. Can. Inst. 43: 319–333 (1969).Google Scholar
  37. 37.
    C.M. Croce, H. Koprowski and H. Eagle, Effect of environmental pH on the efficiency of cellular hybridisation, Proc. Natl. Acad. Sci. U.S.A. 69: 1953–1956 (1972).PubMedCrossRefGoogle Scholar
  38. 38.
    R.L. Davidson and P.S. Gerald, Improved techniques for the induction of mammalian cell hybridization by polyethylene glycol, Somat. Cell Genet. 2: 165–170 (1976).PubMedCrossRefGoogle Scholar
  39. 39.
    S. Brown, F.J. Wiebel, H.V. Gelboin and J.D. Minna, Evidence for linkage between aryl hydrocarbon hydroxylase expression and enzyme markers assigned to human chromosome 2 in human x mouse hybrid cells, in Polycyclic Hydrocarbons and Cancer, vol. 2, pp. 407–415, ed. H.V. Gelboin and P.O.P. Ts’O, Acad. Press, N.Y. (1978).Google Scholar
  40. 40.
    F. Oesch, D.M. Jerina and J.W. Daly, A radiometric assay for hepatic epoxide hydrase activity with 7-3 H styrene oxide, Biochim. Biophys. Acta. 227: 685–691 (1971).PubMedGoogle Scholar
  41. 41.
    J. Singh and F.J. Wiebel, A highly sensitive and rapid fluorometric assay for UDP-glucuronyl transferase using 3-hydroxybenzo[a]-pyrene as substrate, Anal. Biochem. 98: 394–401 (1979).PubMedCrossRefGoogle Scholar
  42. 42.
    O.H. Lowry, N.J. Rosebrough, A.L. Farr and R.J. Randall, Protein measurement with the Folin phenol reagent, J. Biol. Chem. 193: 265–275 (1951).PubMedGoogle Scholar
  43. 43.
    S. Brown, H.K. Oie, A.F. Gazdar, J.D. Minna and U. Francke, Requirement of human chromosomes 19, 6 and possibly 3 for infection of hamster x human hybrid cells with baboon M7 type C virus, Cell 81: 135–143 (1979).CrossRefGoogle Scholar
  44. 44.
    S. Brown, P.A. Lalley and J.D. Minna, Assignment of the gene for peptidase S to chromosome 4 in man and confirmation of peptidase D assignment to chromosome 19, Cytogenet. Cell Genet. 22: 167–171 (1978).PubMedCrossRefGoogle Scholar
  45. 45.
    G. Kellermann, M. Luyten-Kellermann, J.R. Jett, H.L. Moses and R.S. Fontana, Aryl hydrocarbon hydroxylase in man and lung cancer, Human Genet. Suppl., 1: 161–168 (1978).CrossRefGoogle Scholar
  46. 46.
    S. Brown and D.E. Chalmers, Microsomal epoxide hydrolase activity in human x mouse hybrid cells, Biochem. Biophys. Res. Comm. 137: 775–780 (1986).PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Stuart Brown
    • 1
  • Helen Ross
    • 1
  1. 1.Biochemistry DepartmentUniversity of Nottingham Medical SchoolNottinghamEngland

Personalised recommendations