Summary
Phenotypic changes (increased longevity, decreased growth factor requirements, altered cell surface features, growth in semisolid agarose, and SV40 T antigen expression) suggesting in vitro transformation were displayed by human normal colon mucosal epithelial cells transfected with pSV3gpt, a pBR322 recombinant containing the SV40 “early” T antigen coding region and the dominant selectable marker bacterial gene, xanthine-guanine phosphoribosyltransferase. In contrast, control cultures which received neither DNA nor the recombinatn pSV2gpt (which is identical to pSV3gpt but lacks the SV40 T antigen region) were not phenotypically altered.
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References
Boland, C. R.; Montgomery, C. K.; Kim, Y. S. Alterations in human colonic mucin occurring with cellular differentiation and malignant transformation. Proc. Natl. Acad. Sci. 79:2051–2055; 1982.
Boland, C. R.; Montgomery, C. K.; Kim, Y. S. A cancer-associated mucin alteration in benign colonic polyps. Gastroenterol. 82:664–672; 1982.
Boyd, V. A.; Butel, J. Demonstration of infectious deoxyribonucleic acid in transformed cells. I. Recovery of simian virus 40 from yielder and nonyielder transformed cells. J. Virol. 10:399–409; 1972.
Friedman, E. A. Differential response of premalignant epithelial cell classes to phorbol ester tumor promoters and to deoxycholic acid. Cancer Res. 41:4588–4599; 1981.
Friedman, E. A.; Gillin, S.; Lipkin, M. 12-0-Tetradecanoylphorbol-13-acetate stimulation of DNA synthesis in cultured preneoplastic familial polyposis colonic epithelial cells but not in normal colonic epithelial cells. Cancer Res. 44:4078–4086; 1984.
Friedman, E. A.; Higgins, P. J.; Lipkin, M.; Shinya, H.; Gelb, A. M. Tissue culture of human epithelial cells from benign colonic tumors. In Vitro 17:632–644; 1981.
Friedman, E. A.; Steinberg, M. Disrupted communication between late-stage premalignant human colon epithelial cells by 12-O-Tetradecanolyphorbol-13-acetate. Cancer Res. 42:5096–5105; 1982.
Friedman, E. A.; Urmacher, C.; Winawer, S. A model for human colon carcinoma evolution based on the differential response of culture preneoplastic, premalignant, and malignant cells to 12-O-Tetradecanoylphorbol-13-acetate. Cancer Res. 44:1568–1578; 1984.
Friedman, E. A.; Verderame, M.; Winawer, S.; Pollack, R. Actin cytoskeletal organization loss in the benign-to-malignant tumor transition in cultured human colonic epithelial cells. Cancer Res. 44:3040–3050; 1984.
Graham, G. L.; van der Eb, A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467; 1973.
Itzkowitz, S. H.; Kim, Y. S. New carbohydrate tumor markers. Gastroenterol. 90:491–494; 1986.
Lipotich, G.; Moyer, M. P.; Moyer, R. C. Rescue of SV40 following transfection of TC7 cells with cellular DNAs containing complete and partial SV40 genomes. Mol. Gen. Genet. 186:78–81; 1982.
MacPherson, I. A.; Montagnier, L. Agar suspension culture for the selective assay of cells transformed by polyoma virus. Virology. 23:291–294; 1964.
Maniatis, T.; Fritsch, E. F.; Sambrook, J. Molecular cloning, a laboratory manual. New York: Cold Spring Harbor Laboratory Press; 1982.
McCutchan, J. H.; Pagano, J. S. Enhancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J. Natl. Cancer Inst. 41:351–357; 1968.
Moyer, M. P. Culture of human gastrointestinal epithelial cells. Proc. Soc. Exp. Biol. Med. 174:12–15; 1983.
Moyer, M. P. A rapid, reproducible method for processing human solid tumors for in vitro culture. J. Tiss. Cult. Meth. 8:63–68; 1983.
Moyer, M. P.; Aust, J. B. Human colon cells: Culture and in vitro transformation. Science 224:1445–1447; 1984.
Moyer, M. P.; Moyer, R. C. Virus-induced transformation. In: Cameron, I. and Pool, T. B., eds. The transformed cell. New York: Academic Press; 1981:189.
Moyer, M. P.; Aust, J. B.; Dixon, P. S.; Levine, B. A.; Sirinek, K. R. Glucagon enhances growth of cultured human colon cancer cells. Am. J. Surg. 150:676–679; 1985.
Moyer, M. P.; Dixon, P.; Escobar, D.; Aust, J. B. Gastrointestinal cells: Growth factors, transformation, and malignancy. In: Skehan, P. and Friedman, S. J., eds. Growth, cancer, and the cell cycle. Clifton, New Jersey: Humana Press, Inc.; 1984:297–305.
Moyer, M. P.; Page, C. P.; Moyer, R. C. In vitro culture of gastrointestinal epithelial cells and tissues. In: Webber, M. M. and Sekely, L. I., eds. In vitro models for cancer research vol. 1, Boca Raton: CRC Press Inc., Fla. 1985:183–205.
Mulligan, R. C.; Berg, P. Selection for animal cells that express theEscherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc. Natl. Acad. Sci. 78:2072–2076; 1981.
Rutzky, L. P. The biology of human colon tumor cells in culture. In: Advances in cell culture. vol. 4. New York: Academic Press, 1985:47–83.
Shamsuddin, A. K. M.; Weiss, L.; Phelps, P. C.; Trump, F. B. Colon epithelium. IV. Human colon carcinogenesis. Changes in human colon mucosa adjacent to and remote from carcinomas of the colon. J. Natl. Cancer Inst. 66:413–419; 1981.
Weinberg, R. A. The action of oncogenes in the cytoplasm and nucleus. Science 230:770–776; 1985.
Yoakum, G. H. Protoplast-fusion: A method to transfect human cells for gene isolation, oncogene testing and construction of specialized cell lines. BioTechniques. Jan–Feb:24–30; 1984.
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Moyer, M.P., Bradley Aust, J. Phenotypic changes and gene expression in human colon mucosal epithelial cells upon transfection of a SV40 DNA-GPT recombinant. In Vitro Cell Dev Biol 23, 141–146 (1987). https://doi.org/10.1007/BF02623595
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DOI: https://doi.org/10.1007/BF02623595