Summary
The tumor promoter phorbol 12,13-didecanoate (PDD) significantly altered the growth properties of early passage normal human skin cells in vitro in culture medium supplemented with elevated concentrations of selected amino acids. Continuous treatment of cell with 10−7 or 10−8 M PDD resulted in a 5 to 10-fold increase in saturation density at early passages followed by a long-term two- to fourfold increase. The PDD-treated cultures remained in exponential growth at cell densities greater than 10-fold higher than the control cultures. Removal of PDD from the culture medium while the cells were at a high cell density resulted in a return to near-normal saturation density by the subsequent passage. Anchorage independent growth of normal human cells in methylcellulose was also promoted by PDD in a dose dependent manner, with prior subculturing in the presence of PDD being required for maximal colony formation. The structural analog 4α-phorbol 12,13-didecanoate failed to elicit similar cellular responses.
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References
Berenblum, I. Sequential aspects of chemical carcinogensis: Skin. Becker, F. F. ed. Cancer: A comprehensive treatise. Vol. 1. New York: Plenum Publishing Corp.; 1975: 323–344.
Blumberg, P. M.; Driedger P. E.; Rossow, P. W. Effect of a phorbol ester on transformation-sensitive surface protein of chick fibroblasts. Nature 264: 446–447; 1976.
Colburn, N. H.; Former, B. F.; Nelson, K. A.; Yuspa, S. H. Tumor promoter induces anchorage independence irreversibly. Nature 281: 589–591; 1979.
Fisher, P. B.; Bozzone, J. H.; Weinstein, I. B. Tumor promoters and epidermal growth factors stimulate anchorage-independent growth of adenovirus-transformed rat embryo cells. Cell 18: 695–705; 1979.
Ishii, D. N.; Fibach, E.; Yamasaki, H.; Weinstein, I. B. Tumor promoters inhibit morphological differentiation in cultured mouse neuroblastoma cells. Science 200: 556–559; 1978.
Mondal, S.; Brankow, D. W. Heidelberger, C. Two-stage chemical oncogenesis in cultures of C3H/10T1/2 cells. Cancer Res. 36: 2254–2260; 1976.
Rovera, G.; O’Brien, T. G.; Diamond L. Tumor promoters inhibit spontaneous differentiation of Friend erythroleukemia cells in culture. Proc. Natl. Acad. Sci. USA 74: 2894–2898; 1977.
Weinstein, I. B.; Lee, L-S.; Fisher, P. B.; Mufson, A.; Yamasaki, H. Action of phorbol esters in cell culture: Mimicry of transformation, altered differentiation, and effects on cell membranes. J. Supramol. Struct. 12: 195–208; 1979.
Diamond, L.; O’Brien, S.; Donaldson, C.; Shimazu, Y. Growth stimulation of human diploid fibroblasts by the tumor promoter 12-O-tetradecanoyl phorbol-13-acetate. Int. J. Cancer 13: 721–730; 1974.
Gansler, T.; Kopelovich, L. Effects of 12-O-tetradecanoyl phorbol-13-acetate and epidermal growth factor on the proliferation of human mutant fibroblastsin vitro. Cancer Lett. 13: 315–323; 1981.
Kinzel, V.; Richards, J.; Stöhr, M. Early effects of the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate on the cell cycle traverse of asynchronous HeLa cells. Cancer Res. 41: 300–305; 1981.
Kopelovich, L.; Bias, N. E.; Helson, L. Tumor promoter alone induces neoplastic transformation of fibroblasts from humans genetically predisposed to cancer. Nature 282: 619–621; 1979.
Liebermann, D.; Hoffman-Liebermann, B.; Sachs, L. Regulation of gene expression by tumor promoters. II. Control of cell shape and developmental programs for macrophages and granulocytes in human myeloid leukemia cells. Int. J. Cancer 28: 285–291; 1981.
Rovera, G.; Santoli, D.; Damsky, C. Human promyelocytic leukemia cells in culture differentiate into macrophage-like cells when treated with a phorbol diester. Proc. Natl. Acad. Sci. USA 76: 2779–2783; 1979.
Milo, G. E.; Oldham, J. W.; Zimmerman, R.; Hatch, G. G.; Weisbrode, S. A. Characterization of human cells transformed by chemical and physical carcinogensin vitro. In Vitro 17: 719–729; 1981.
Riegner, D. A.; McMichael, T.; Berno, J. C.; Milo, G. E. Processing of human tissue to establish primary cultures in vitro. TCA Manual 2: 273–275; 1976.
Milo, G. E.; Ackerman, G. A.; Noyes, I. Growth and ultrastructural characterization of proliferating human keratinocytes in vitro without added extrinsic factors. In Vitro 16: 20–30; 1980.
Tomei, L. D.; Cheney, J. C.; Wenner, C. E. The effect of phorbol esters on the proliferation of C3H-10T1/2 mouse fibroblasts: Consideration of both stimulatory and inhibitory effects. J. Cell. Physiol. 107: 385–389; 1981.
Antecol, M. H.; Mukherjee, B. B. Effects of 12-O-tetradecanoyl phorbol-13-acetate on fibroblasts from individuals genetically predisposed to cancer. Cancer Res. 42: 3870–3879; 1982.
Fisher, P. B.; Miranda, A. F.; Mufson, R. A.; Weinstein, I. B. Effects of teleocidin and the phorbol ester tumor promoters on cell transformation, differentiation, and phospholipid metabolsim. Cancer Res. 42: 2829–2835; 1982.
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This work was supported by grants from the Air Force Office of Scientific Research, Department of Defense (AFOSR-80-0283) and the National Cancer Institute, Department of Health and Human Services (P-30-CA-16058).
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Trewyn, R.W., Gatz, H.B. Altered growth properties of normal human cells induced by phorbol 12,13-didecanoate. In Vitro 20, 409–415 (1984). https://doi.org/10.1007/BF02619587
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DOI: https://doi.org/10.1007/BF02619587