Summary
Susceptibility to chemically induced transformation changed as a rat embryo cell culture was passaged. For the first 35 to 60 passages, the cultures were diploid and resistant to transformation by chemical carcinogens. However, cultures infected with a murine leukemia virus were transformed by chemicals. For the next 60 passages, the cultures were heteroploid, but retained contact inhibition and were not tumorigenic. Even without addition of heterotypic viruses, these heteroploid cultures could be transformed by chemicals, but the endogenous rat C-type virus could be demonstrated in the transformed cultures. At higher passages, the rates of spontaneous transformation gradually increased so that the cultures could not be used for transformation studies.
Chemically induced transformation of the stable heteroploid cell line (F1706) was manifested by an easy to read focal alteration. Initial observations based on these foci were confirmed by inoculating the morphologically altered cells into isogeneic newborn rats.
A number of carcinogenic and noncarcinogenic chemical analogues were tested for their ability to transform F1706 cultures. The compounds tested included 4 azo dyes, 12 polycyclic hydrocarbons, 12 aromatic amines, and 7 miscellaneous compounds. Based on the known activities of the same chemicals in rodents, certain active compounds failed to induce transformation in any test, and others induced transformation in only some tests, but these in vitro tests, if used as a screening assay, would have been correct in 82% of all individual tests, and over-all, would have correctly predicted the carcinogenic activity of 33 of the 35 agents tested.
Similar content being viewed by others
References
Earle, W. R. 1943. Production of malignancy in vitro. IV. The mouse fibroblast cultures and changes seen in the living cells. J. Natl. Cancer Inst. 4: 165–212.
Berwald, Y., and L. Sachs. 1963. In vitro cell transformation with chemical carcinogens. Nature 200: 1182–1184.
Borenfreund, E., M. Krim, F. K. Sanders, S. Sternberg, and A. Bendich. 1966. Malignant conversion of cells in vitro by carcinogens and viruses. Proc. Natl. Acad. Sci. U.S.A. 56: 672–679.
DiPaolo, J. A., and P. J. Donovan. 1967. Properties of Syrian hamster cells transformed in the presence of carcinogenic hydrocarbons. Exp. Cell Res. 48: 361–377.
Heidelberger, C., and P. T. Iype. 1967. Malignant transformation in vitro by carcinogenic hydrocarbons. Science 155: 214–217.
Sanders, F. K., and B. O. Burford. 1967. Morphological conversion of cells in vitro by N-nitrosomethylurea. Nature 213: 1171–1173.
DiPaolo, J. A., A. S. Rabson, and R. A. Malmgren. 1968. In vitro viral transformation cells. J. Natl. Cancer Inst. 40: 757–770.
Huberman, E., S. Salzberg, and L. Sachs. 1968. The in vitro induction of an increase in cell multiplication and cellular life span by the water-soluble carcinogen dimethylnitrosamine. Proc. Natl. Acad. Sci. U.S.A. 59: 77–82.
Kuroki, T., and H. Sato 1968. Transformation and neoplastic development in vitro of hamster embryonic cells by 4-nitroquinoline-1-oxide and its derivatives. J. Natl. Cancer Inst. 41: 53–71.
Sanders, F. K., and B. O. Burford. 1968. Morphological conversion, hyperconversion and reversion of mammalian cells treated in vitro with N-nitrosomethylurea. Nature 220: 448–453.
Sivak, A., and B. L. Van Duuren. 1968. Studies with carcinogens and tumor-promoting agents in cell culture. Exp. Cell Res. 49: 572–583.
Chen, T. T., and C. Heidelberger. 1969. Quantitative studies on the malignant transformation of mouse prostate cells by carcinogenic hydrocarbons in vitro. Int. J. Cancer 4: 166–178.
DiPaolo, J. A., R. L. Nelson, and P. J. Donovan. 1969. Sarcoma-producing cell lines derived from clones transformed in vitro by benzo (a) pyrene. Science 165: 917–918.
DiPaolo, J. A., P. Donovan, and R. Nelson. 1969. Quantitative studies of in vitro transformation by chemical carcinogens. J. Natl. Cancer Inst. 42: 867–876.
Namba, M., H. Masuji, and J. Sato. 1969. Carcinogenesis in tissue cultures. IX. Malignant transformation of cultured rat cells treated with 4-nitroquinoline-1-oxide. Jap. J. Exp. Med. 39: 253–265.
Takaki, R., M. Takii, and T. Ikegami. 1969. Preliminary studies on the in vitro carcinogenesis of rat thymus cells by N-methyl-N’-nitro-N-nitrosoguanidine. Gann 60: 661–662.
Mondal, S., and C. Heidelberger. 1970. In vitro malignant transformation by methylcholanthrene of the progeny of single cells derived from C3H mouse prostate. Proc. Natl. Acad. Sci. U.S.A. 65: 219–225.
Freeman, A. E., P. J. Price, H. J. Igel, J. C. Young, J. M. Maryak, and R. J. Huebner. 1970. Morphological transformation of rat embryo cells induced by diethylnitrosamine and murine leukemia viruses. J. Natl. Cancer Inst. 44: 65–78.
Freeman, A. E., P. J. Price, R. J. Bryan, R. J. Gordon, R. V. Gilden, G. J. Kelloff, and R. J. Huebner. 1971. Transformation of rat and hamster embryo cells by extracts of city smog. Prog. Natl. Acad. Sci U.S.A. 68: 445–449.
Grover, P. L., P. Sims, E. Huberman, H. Marquardt, T. Kuroki, and C. Heidelberger. 1971. In vitro transformation of rodent cells by K-region derivatives of polycyclic hydrocarbons. Proc. Natl. Acad. Sci. U.S.A. 68: 1098–1101.
Price, P. J., A. E. Freeman, W. T. Lane, and R. J. Huebner. 1971 Morphological transformation of rat embryo cells by the combined action of 3-methylcholanthrene and Rauscher leukaemia virus. Nature New Biol. 230: 144–146.
Rhim, J. S., W. Vass, H. Y. Cho, and R. J. Huebner. 1971. Malignant transformation induced by 7,12-dimethylbenz(a)anthracene in rat embryo cells infected with Rauscher leukemia virus. Int. J. Cancer 7: 65–74.
Rhim, J. S., B. Creasy, and R. J. Huebner. 1971. Production of altered cell foci by 3-methylcholanthrene in mouse cells infected with AKR leukemia virus. Proc. Natl. Acad. Sci. U.S.A. 68: 2212–2216.
DiPaolo, J. A., R. L. Nelson, and P. J. Donovan. 1972. In vitro transformation of Syrian hamster embryo cells by diverse chemical carcinogens. Nature 235: 278–280.
Huberman, E., P. J. Donovan, and J. A. DiPaolo. 1972. Mutation and transformation of cultured mammalian cells by N-acetoxy-N-2-fluorenylacetamide. Proc. Natl. Cancer Inst. 48: 837–840.
Mishra, N. K., and W. L. Ryan. 1973. Effect of 3-methylcholanthrene and dimethylnitrosamine on chronically infected with Rauscher leukemia virus. Int. J. Cancer 11: 123–130.
Freeman, A. E., P. J. Price, E. M. Zimmerman, G. J. Kelloff, and R. J. Huebner. 1973. RNA tumor virus genomes as determinants of chemically-induced transformation in vitro. In: R. M. Dutcher and L. Chieco-Bianchi (Eds.),Unifying Concepts of Leukemia. Bibl. Haemat., No. 39. Karger, Basel, pp. 617–634.
Rhim, J. S., and R. J. Huebner, 1973. Dual effect of RNA tumor viruses and carcinogenic chemicals or DNA viruses. In: R. M. Dutcher and L. Chieco-Bianchi (Eds.),Unifying Concepts of Leukemia. Bibl. Haemat., No. 39. Karger, Basel, pp. 635–648.
Williams, G. M., J. M. Elliott, and J. H. Weisburger. 1973. Carcinoma after malignant conversion in vitro of epithelial-like cells from rat liver following exposure to chemical carcinogens. Cancer Res. 33: 606–612.
Freeman, A. E., E. K. Weisburger, J. H. Weisburger, R. G. Wolford, J. M. Maryak, and R. J. Huebner. 1973. Transformation of cell cultures as an indication of the carcinogenic potential of chemicals. J. Natl. Cancer Inst. 51: 799–808.
Freeman, A. E., R. V. Gilden, M. L. Vernon, R. G. Wolford, P. E. Hugunin, and R. J. Huebner. 1973. 5-bromo-2′-deoxyuridine potentiation of transformation of rat-embryo cells induced in vitro by 3-methylcholanthrene: induction of rat leukemia virus gas antigen in transformed cells. Proc. Natl. Acad. Sci. U.S.A. 70: 2415–2419.
Hayflick, L., and P. S. Moorhead. 1961. The serial cultivation of human diploid cell strains. Exp. Cell Res. 25: 585–621.
Hayflick, L. 1970. Aging under glass. Exp. Gerontol. 5: 291–303.
Zimmerman, E. M., R. E. Kouri, K. Higuchi, F. Laird, and A. E. Freeman 1975. Uptake, metabolism, and persistence of 3-methylcholanthrene in rat embryo cells infected with murine leukemia virus. Cancer Res. 35: 139–143.
Igel, H. J., R. J. Huebner, H. C. Turner, P. Kotin, and H. L. Falk 1969. Mouse leukemia virus activation by chemical carcinogens. Science 166: 1624–1626.
Freeman, A. E., G. J. Kelloff, R. V. Gilden, W. T. Lane, A. P. Swain, and R. J. Huebner. 1971. Activation and isolation of hamster specific C-type RNA viruses from tumors induced by cell cultures transformed by chemical carcinogens. Proc. Natl. Acad. Sci. U.S.A. 68: 2386–2390.
Freeman, A. E., G. J. Kelloff, M. L. Vernon, W. T. Lane, W. I. Capps, S. D. Bumgarner, H. C. Turner, and R. J. Huebner. 1974. Prevalence of endogenous type-C virus in normal hamster tissues and hamster tumors induced by chemical carcinogens, Simian Virus 40, and polyoma virus. J. Natl. Cancer Inst. 52: 1469–1476.
Author information
Authors and Affiliations
Additional information
Supported in part under contract NIH-NCI-70-2068 within the Special Virus-Cancer Program of the National Cancer Institute, NIH, PHS.
Rights and permissions
About this article
Cite this article
Freeman, A.E., Igel, H.J. & Price, P.J. Carcinogenesis in vitro. In Vitro 11, 107–116 (1975). https://doi.org/10.1007/BF02624083
Issue Date:
DOI: https://doi.org/10.1007/BF02624083