High frequency of carcinogen-induced early, preneoplastic changes in rat tracheal epithelial cells in culture.

To study the mechanisms of carcinogenesis, we have developed a system that uses normal cells from an environmentally and epidemiologically relevant tissue, respiratory epithelium. The induction of preneoplastic variants of epithelial cells in culture was quantitated on a per-cell basis following exposure of rat tracheal epithelial (RTE) cells in vitro to the direct-acting carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Following treatment of normal RTE cells, large colonies of altered cells exhibiting an enhanced growth potential under selective culture conditions were observed, while normal RTE cells ceased proliferation after several cell doublings. After further growth in culture, these altered cells acquired the ability to grow in semisolid medium and to produce squamous cell carcinomas when injected into nude mice. The induction of enhanced growth variants of RTE cells by MNNG occurred with a high frequency (greater than or equal to 2.6%/colony-forming cell). In addition, a linear dose-response curve with a slope of approximately 1 was observed when the logarithm of MNNG-induced transformation frequency was plotted versus the logarithm of MNNG dose. These results are consistent with a one-hit mechanism for induction of preneoplastic variants of RTE cells by MNNG. Similar frequencies and kinetics of induction of preneoplastic variants in other culture systems using diploid cells have been observed, suggesting a common mechanism for this early step in carcinogenesis. The RTE cell system will be useful for mechanistic studies of early as well as late changes in the development of neoplasia by epithelial cells.

[1]  T. Hesterberg,et al.  ROLE OF GENE AND CHROMOSOMAL MUTATIONS IN CELL TRANSFORMATION , 1983, Annals of the New York Academy of Sciences.

[2]  P. Nettesheim,et al.  Localization of specific lesions in dimethylbenz(a)anthracene-preexposed tracheal explants. , 1982, The American journal of pathology.

[3]  R. Demars,et al.  Clonal analysis of the stepwise appearance of anchorage independence and tumorigenicity in CAK, a permanent line of mouse cells. , 1982, Cancer research.

[4]  M. O. Bradley,et al.  Neoplastic Transformation of Syrian Hamster Epid , 1982 .

[5]  A. Pardee,et al.  Growth of Cells in Hormonally Defined Media , 1982 .

[6]  T. Nomura.  Parental exposure to X rays and chemicals induces heritable tumours and anomalies in mice. , 1982, Nature.

[7]  S. Yuspa,et al.  Mouse skin cells resistant to terminal differentiation associated with initiation of carcinogenesis , 1981, Nature.

[8]  R. Tennant,et al.  In vitro transformation of Syrian hamster epidermal cells by N-methyl-N'-nitro-N-nitrosoguanidine. , 1981, Cancer research.

[9]  E. Miller,et al.  Mechanisms of chemical carcinogenesis , 1981, Cancer.

[10]  J. Barrett,et al.  Inhibition of terminal differentiation of hamster epidermal cells in culture by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate. , 1981, Cancer research.

[11]  J. Cairns.  The origin of human cancers , 1981, Nature.

[12]  M. Kulesz-Martin,et al.  Quantitative assay for carcinogen altered differentiation in mouse epidermal cells. , 1980, Carcinogenesis.

[13]  C. Heidelberger,et al.  Probabilistic view of the transformation of cultured C3H/10T1/2 mouse embryo fibroblasts by 3-methylcholanthrene. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Little,et al.  Relationship between x-ray exposure and malignant transformation in C3H 10T1/2 cells. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J. Gart,et al.  Mathematical models and the statistical analyses of cell transformation experiments. , 1979, Cancer research.

[16]  P. Nettesheim,et al.  Dynamics of neoplastic development in carcinogen-exposed tracheal mucosa. , 1979, Cancer research.

[17]  P. Nettesheim,et al.  Repopulation of denuded tracheal grafts with normal, preneoplastic, and neoplastic epithelial cell populations. , 1978, Cancer research.

[18]  C. Wigley.  Neoplastic transformation in differentiated epithelial cell systems in vitro , 1978 .

[19]  P. Nettesheim,et al.  In vitro development of oncogenicity in cell lines established from tracheal epithelium preexposed in vivo to 7,12-dimethylbenz(a)anthracene. , 1978, Cancer research.

[20]  J. Barrett,et al.  Relationship between somatic mutation and neoplastic transformation. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[21]  P. Nettesheim,et al.  Increased in vitro growth capacity of tracheal epithelium exposed in vivo to 7, 12-dimethylbenz(a)anthracene. , 1977, Cancer research.

[22]  H. Green,et al.  Seria cultivation of strains of human epidemal keratinocytes: the formation keratinizin colonies from single cell is , 1975, Cell.

[23]  C. Harris,et al.  Altered differentiation of mouse epidermal cells treated with retinyl acetate in vitro. , 1974, Experimental cell research.

[24]  J. Barrett.  Use of quantitative cell transformation assays in risk estimation , 1985 .

[25]  P. Nettesheim,et al.  Neoplastic transformation of primary tracheal epithelial cell cultures. , 1983, Carcinogenesis.

[26]  M. O. Bradley,et al.  Neoplastic transformation of Syrian hamster epidermal cells in vitro. , 1982, Cancer research.

[27]  J. Rheinwald.  Serial cultivation of normal human epidermal keratinocytes. , 1980, Methods in cell biology.

[28]  J. Rheinwald.  Chapter 15 Serial Cultivation of Normal Human Epidermal Keratinocytes , 1980 .

[29]  H Green,et al.  Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. , 1975, Cell.

[30]  C. Heidelberger.  Chemical oncogenesis in culture. , 1973, Advances in cancer research.