Summary
Human fibroblasts (KMST-6) immortalized by treatment with60Co gamma rays were further neoplastically transformed by transfection of the c-Ha-ras oncogene from human lung cancer. The ras-transfected cells formed undifferentiated fibrosarcoma in nude mice. One of the tumors was recultured and a neoplastic human fibroblast line, KMST-6/RAS, was established. To analyze multistep carcinogenesis of human cells, the cellular characteristics of these genetically matched immortalized (KMST-6) and neoplastic (KMST-6/RAS) cell lines were studied in detail. KMST-6/RAS cells showed an increased saturation density, colony formation on confluent monolayers of normal human fibroblasts, proliferation in neomycin-containing medium, anchorage-independent growth, and enhanced expression of the transfected c-Ha-ras oncogene, whereas the immortalized cells did not demonstate these characteristics. Unexpectedly, growth of KMST-6/RAS cells was serum-dependent, although they were neoplastic. Interestingly, the neoplastic cells did not show the criss-crossing or piling up growth pattern characteristic of transformed rodent fibroblasts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bookland, E. A.; Swaminathan, S.; Oyasu, R., et al. Tumorigenic transformation and neoplastic progression of human uroepithelial cells after exposure in vitro to 4-aminobiphenyl or its metabolites. Cancer Res. 52:1606–1614; 1992.
Chen, T. R. In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Exp. Cell Res. 104:225–262; 1977.
Chomczynski, P.; Sacchi, N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162:156–159; 1987.
DiPaolo, J. A.; Woodworth, C. D.; Popescu, N. C., et al. Induction of human cervical squamous cell carcinoma by sequential transfection with human papillomavirus 16 DNA and viral Harvey ras. Oncogene 4:395–399; 1989.
Dürst, M.; Gissmann, L.; Ikenberg, H., et al. A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions. Proc. Natl. Acad. Sci. USA 80:3812–3815; 1983.
Freund, R.; Bronson, R. T.; Benjamin, T. L. Separation of immortalization from tumor induction with polyoma large T mutants that fail to bind the retinoblastoma gene product. Oncogene 7:1979–1987; 1992.
Gerwin, B. I.; Spillare, E.; Forrester, K., et al. Mutant p53 can induce tumorigenic conversion of human bronchial epithelial cells and reduce their responsiveness to a negative growth factor, transforming growth factor β1. Proc. Natl. Acad. Sci. USA 89:2759–2763; 1992.
Hurlin, P. J.; Maher, V. M.; McCormick, J. J. Malignant transformation of human fibroblasts caused by expression of a transfected T24 HRAS oncogene. Proc. Natl. Acad. Sci. USA 86:187–191; 1989.
Hurlin, P. J.; Dennis, G. F.; Veronica, V. M., et al. Morphological transformation, focus formation, and anchorage independence induced in diploid human fibroblasts by expression of a transfected H-ras oncogene. Cancer Res. 47:5752–5757; 1987.
McCormick, J. J.; Maher, V. M. Towards an understanding of the malignant transformation of diploid human fibroblasts. Mutat. Res. 199:273–291; 1988.
Mihara, K.; Bai, L.; Kano, Y., et al. Malignant transformation of human fibroblasts previously immortalized with60Co gamma rays. Int. J. Cancer 50:639–643; 1992.
Namba, M.; Nishitani, K.; Fukushima, F., et al. Multistep process of neoplastic transformation of normal human fibroblasts by60Co gamma rays and Harvey sarcoma viruses. Int. J. Cancer 37:419–423; 1986.
Namba, M.; Nishitani, K.; Fukushima, F., et al. Multi-step neoplastic transformation of normal human fibroblasts by C-60 gamma rays and Ha-ras oncogenes. Mutat. Res. 199:415–423; 1988.
Namba, M.; Nishitani, K.; Hyodoh, F., et al. Neoplastic transformation of human diploid fibroblasts (KMST-6) by treatment with60Co gamma rays. Int. J. Cancer 35:275–280; 1985.
Newbold, R. F.; Overell, R. W. Fibroblast immortality is a prerequisite for transformation by EJ c-Ha-ras oncogene. Nature 304:648–651; 1983.
Pfeifer, A. M. A.; Jones, R. T.; Bowden, P. E., et al. Human bronchial epithelial cells transformed by the c-raf and c-myc protooncogenes induce multidifferentiated carcinomas in nude mice: a model for lung carcinogenesis. Cancer Res. 51:3793–3801; 1991.
Rhim, J. S.; Yoo, J. H.; Park, J. H., et al. Evidence for the multistep nature of in vitro human epithelial cell carcinogenesis. Cancer Res. 50(Suppl):5653s-5657s; 1990.
Sager, R.; Tanaka, K.; Lau, C. C., et al. Resistance of human cells to tumorigenesis induced by cloned transforming genes. Proc. Natl. Acad. Sci. USA 80:7601–7605; 1983.
Schaeffer, W. I. Usage of vertebrate, invertebrate and plant cell, tissue and organ culture terminology. In Vitro 20:19–23; 1983.
Smith, S. H.; Owens, R. B.; Hiller, A. J., et al. The biology of human cells in tissue culture. I. Characterization of cells derived from osteogenic sarcomas. Int. J. Cancer 17:219–234; 1976.
Thraves, P.; Salehi, Z.; Dritschilo, A., et al. Neoplastic transformation of immortalized human epidermal keratinocytes by ionizing radiation. Proc. Natl. Acad. Sci. USA 87:1174–1177; 1990.
Wilson, D. M.; Yang, D.; Dillberger, J. E., et al. Malignant transformation of human fibroblasts by a transfected N-ras oncogene. Cancer Res. 50:5587–5593; 1990.
Yang, D.; Louden, C.; Reinhold, D. S., et al. Malignant transformation of human fibroblast cell strain MSU-1.1 by (+)-7β, 8α-dihydroxy-9α, 10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Proc. Natl. Acad. Sci. USA 89:2237–2241; 1992.
Yuasa, Y.; Shrivastava, S. K.; Dunn, Y. C., et al. Acquisition of transformaing properties by alternative point mutations within c-bas/has human proto-oncogene. Nature 303:775–779; 1983.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Jahan, I., Mihara, K., Bai, L. et al. Neoplastic transformation and characterization of human fibroblasts by treatment with60Co gamma rays and the human c-Ha-ras oncogene. In Vitro Cell Dev Biol - Animal 29, 763–767 (1993). https://doi.org/10.1007/BF02634342
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02634342