Summary
Several different classes of chemical carcinogens induced the transformation of human fibroblasts grown in vitro. Characteristics of the events that occur from time of treatment through the expression of neoplastic transformation are presented. The S-phase appeared to be the portion of the cell cycle most vulnerable to insult. Staging of the cells by blocking them in G1 before releasing them to proceed through scheduled DNA synthesis (S) was required to induce reproducible transformation. Compounds such as insulin were added to the cells upon release from the block to sensitize the cells to the carcinogen that was added during S. Growth of the transformed cells as distinct from nontransformed cells was promoted by growth in medium supplemented with 8X nonessential amino acids. Carcinogen-treated cells in the early stage of transformation exhibited abnormal colony morphology and were able to grow at 41°C, in air atmosphere, and in medium supplemented with only 1% serum. In addition, the transformed cells were insensitive to KB cell lysate and exhibited density independent, as well as anchorage independent, growth (i.e., growth in 0.33% agar). Cells that grew in soft agar also produced undifferentiated mesenchymal tumors in preirradiated nude mice.
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Milo, G.; DiPaolo, J. Neoplastic transformation of human diploid cellsin vitro after chemical carcinogen treatment. Nature 275: 130–132; 1978.
Milo, G.; Olsen, R.; Weisbrode, S.; McCloskey, J. Feline sarcoma virus-induced in vitro progression from premalignant to neoplastic transformation of human diploid cells. In Vitro 16: 813–822; 1980.
Milo, G.; Weisbrode, S.; Zimmerman, R.; McCloskey, J. Ultraviolet radiation induced transformation of normal human cellsin vitro. Chem. Biol. Interact. 36: 45–59; 1981.
Milo, G.; DiPaolo, J. Presentization of human cells with extrinsic signals to induced chemical carcinogenesis. Int. J. Cancer. 26: 805–812; 1981.
Reigner, D.; McMichael, T.; Berno, J.; Milo, G. Processing of human tissue to establish primary cultures in vitro. TCA Manual 2: 273–276; 1976.
Oldham, J.; Allred, L.; Milo, G.; Kindig, O.; Capen, C. The toxicological evaluation of the mycotoxins T-2 and T-2 tetraol using normal human fibroblastsin vitro. Toxicol. Appl. Pharmacol. 52: 159–168; 1980.
Van-Nest, G.; Grimes, W. Conconavalin A-induced agglutination and tumorigenicity in virally and spontaneously transformed cells derived from BALB/C mice. Cancer Res. 34: 1408–1412; 1974.
Temin, H. Control of cell multiplication of uninfected chicken cells and chicken cells converted by avian sarcoma viruses. J. Cell Physiol. 74: 9–12; 1969.
Oldham, J.; Milo, G.; Kadlubar, F. Effect of acidic pH on thein vitro transformation of normal human fibroblasts byN-OH arylamines (abstr.) Soc. of Toxicol. 19th Annual Meeting; A-37; 1980.
Casto, B.; Hatch, G. Developments in neoplastic transformation. Berky, J.; Sherrod, C. eds.In Vitro toxicity testing. Philadelphia, PA: Franklin Institute Press; 1977; 192–214.
Ekelman, K.; Milo, G. Cellular uptake, transport and macromolecular binding of benzo(a)pyrene and 7,12-dimethylbenz(a)anthracene by human cellsin vitro. Cancer Res. 38: 3026–3032; 1978.
Kadlubar, F.; Miller, J.; Miller, E. GuanylO 6-arylamination andO 6-arylation of DNA by the carcinogenN-hydroxy-1-naphthylamine. Cancer Res. 38: 3628–3638; 1978.
Kakunaga, T. Neoplastic transformation of human diploid fibroblast cells by chemical carcinogens. Proc. Natl. Acad. Sci. USA 75: 1334–1338; 1978.
Borke, G. X-ray-inducedin vitro neoplastic transformation of human diploid cells. Nature 288: 776–778; 1980.
Turk, B.; Milo, G. Anin vitro study of senescent events in human embryonic (WI-38) cells: 1. Change in enzyme activities of cellular and membrane-associated enzymes of untreated and cortisone acetate treated cultures during senescence. Arch. Biochem. Biophys. 16: 46–53; 1974.
McHale, J. S.; Moulton, M.; McHale, J. T. Limited culture life span of human diploid cells as a function of metabolic time instead of division potential. Exp. Gerontol. 6: 89–93; 1971.
Cristofalo, V. Animal cell cultures as a model system for the study of aging. Adv. Gerontological Res. 4: 45–76; 1971.
Grisham, J.; Greenburg, D.; Smith, G.; Kaufman, D., Temporary culture in isoleucine-free medium enhances transformation of 10 T1/2 cells byN-methyl-N′-nitrosoguanidine (MNNG). Biochem. Biophys. Res. Commun. 87: 969–975; 1979.
Hatch, G.; Balwierz, P.; Casto, B.; DiPaolo, J. Characteristics of hamster cells transformed by the combined action of chemical and virus Int. J. Cancer 21: 121–127; 1978
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This work was supported in part by National Cancer Institute Grant ROI-CA-25907 and Air Force Office of Scientific Research Grant F49620-77-C0110 and EPA-R806638. The hydrazine compounds were furnished by Ms. Marilyn George and Dr. Kenneth Back, AFSOR Toxicology Division, Wright Patterson Air Force Base, Dayton, OH. The hydroxylate and phenyl napthylamines were furnished by Dr. Fred Kadlubar, Division of Chemical Carcinogenesis at the National Center for Toxicological Research, Jefferson, AR.
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Milo, G.E., Oldham, J.W., Zimmerman, R. et al. Characterization of human cells transformed by chemical and physical carcinogens in vitro. In Vitro 17, 719–729 (1981). https://doi.org/10.1007/BF02628409
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DOI: https://doi.org/10.1007/BF02628409