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In Vitro Carcinogenesis with Cell Lines

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In Vitro Toxicity Testing of Environmental Agents

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Abstract

Unlike the Syrian hamster embryo cell system pioneered by Sachs and by DiPaolo, in which chemical carcinogens transform primary or secondary cultures, we and others have used permanent cell lines derived from mouse fibroblasts. As an example, this paper concentrates on the C3H/10T1/2 cells that we developed. These cells are highly susceptible to postconfluence inhibition of cell division, spontaneous transformation is very rare, and they do not produce tumors on inoculation into syngeneic immunosuppressed mice. On treatment with polycyclic hydrocarbons (PAH), X-rays, ultra-violet light, neutrons, some cancer chemotherapeutic drugs, alkylating agents, or tobacco smoke condensate, they lose contact inhibition and form piled-up foci that produce fibrosarcomas in C3H mice. These cells have the cytochrome P448 enzyme system that is required to activate PAH, but lack the capacity to activate other classes of chemical carcinogens. Permanent lines that we recently developed from mouse regenerating liver, when used as irradiated “feeder layers,” activate aflatoxin B1 to transform C3H/10T1/2 cells. In this system: there is no induction of an endogenous retrovirus during chemical transformation; chemically transformed clones have individual tumor-specific and common embryonic cell- surface antigens; and transformation with alkylating agents and antimetabolites is cell-cycle phase-specific. We have obtained two-stage transformation with PAH or UV light as initiators and phorbol esters, anthralin, and saccharin as promoters, and we can now simultaneously study oncogenic transformation and mutagenesis to ouabain resistance. We are using this system to study cellular and molecular mechanisms of chemical oncogenesis, and as a potential prescreen for chemical carcinogens, initiators, and promoters.

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Author information

Authors and Affiliations

  1. Comprehensive Cancer Center, University of Southern California, Los Angeles, California, 90031, USA

    Charles Heidelberger (Director for Basic Research)

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  1. Charles Heidelberger
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Editors and Affiliations

  1. Research Triangle Institute, Research Triangle Park, North Carolina, USA

    Alan R. Kolber , Robert S. DeWoskin  & Thomas J. Hughes ,  & 

  2. National Institute of Environmental Health Sciences (BRAP), Research Triangle Park, North Carolina, USA

    Thomas K. Wong

  3. U.S. Environmental Protection Agency, Research Triangle Park, North Carolina, USA

    Lester D. Grant

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© 1983 Plenum Press, New York

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Heidelberger, C. (1983). In Vitro Carcinogenesis with Cell Lines. In: Kolber, A.R., Wong, T.K., Grant, L.D., DeWoskin, R.S., Hughes, T.J. (eds) In Vitro Toxicity Testing of Environmental Agents. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-3566-5_14

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  • DOI: https://doi.org/10.1007/978-1-4613-3566-5_14

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-3568-9

  • Online ISBN: 978-1-4613-3566-5

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