Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1973 Aug;70(8):2415–2419. doi: 10.1073/pnas.70.8.2415

5-Bromo-2′-deoxyuridine Potentiation of Transformation of Rat-Embryo Cells Induced In Vitro by 3-Methylcholanthrene: Induction of Rat Leukemia Virus gs Antigen in Transformed Cells

Aaron E Freeman *, Raymond V Gilden †, Mina Lee Vernon ‡, Ronald G Wolford ‡, Patricia E Hugunin ‡, Robert J Huebner §
PMCID: PMC433747  PMID: 4134493

Abstract

Low-passage rat-embryo cells were not transformed by 3-methylcholanthrene or by 5-bromo-2′ deoxyuridine. However, prior treatment with bromodeoxyuridine, followed by treatment with methylcholanthrene, resulted in cell transformation about three subpassages after removal of the carcinogen. RNA-directed DNA polymerase activity could not be detected in either normal or transformed cells. However, gs-1 antigen specific for rat C-type RNA virus was detected in cultures derived from bromodeoxyuridine-treated cells. No gs-1 antigen for the C-type RNA virus was detected in cultures that had not been treated with bromodeoxyuridine during the 25 subpassages of these experiments.

High-passage rat-embryo cells, derived from a different cell pool, were transformed by either methylcholanthrene or dimethylbenzanthracene without prior infection with an exogenous virus, and without prior treatment with bromodeoxyuridine, gs-1 antigen for C-type RNA virus was also detected in 4 of 4 randomly selected transformed cell lines; the gs-1 antigen was not detected in any of 4 nontransformed control cultures. Considering these and previously published findings, we conclude that the lowpassage cells cannot be transformed by methylcholanthrene because of powerful cellular controls over the endogenous virus. Bromodeoxyuridine triggers some expressions of the endogenous virus; thus, the bromodeoxyuridine-treated cells are more susceptible to the transforming effects of methylcholanthrene. High-passage rat cells do not maintain perfect control over expression of their endogenous virus; the cell cultures are susceptible to the transforming effects of chemical carcinogens.

Keywords: carcinogen, C-type RNA virus, dimethylbenzanthracene

Full text

PDF
2415

Images in this article

2416Image
on p.2416
2416Image
on p.2416
2418Image
on p.2418
2418Image
on p.2418

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aaronson S. A., Hartley J. W., Todaro G. J. Mouse leukemia virus: "spontaneous" release by mouse embryo cells after long-term in vitro cultivation. Proc Natl Acad Sci U S A. 1969 Sep;64(1):87–94. doi: 10.1073/pnas.64.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aaronson S. A., Todaro G. J., Scolnick E. M. Induction of murine C-type viruses from clonal lines of virus-free BALB-3T3 cells. Science. 1971 Oct 8;174(4005):157–159. doi: 10.1126/science.174.4005.157. [DOI] [PubMed] [Google Scholar]
  3. BERWALD Y., SACHS L. IN VITRO CELL TRANSFORMATION WITH CHEMICAL CARCINOGENS. Nature. 1963 Dec 21;200:1182–1184. doi: 10.1038/2001182a0. [DOI] [PubMed] [Google Scholar]
  4. Berwald Y., Sachs L. In vitro transformation of normal cells to tumor cells by carcinogenic hydrocarbons. J Natl Cancer Inst. 1965 Oct;35(4):641–661. [PubMed] [Google Scholar]
  5. Borenfreund E., Krim M., Sanders F. K., Sternberg S. S., Bendich A. Malignant conversion of cells in vitro by carcinogens and viruses. Proc Natl Acad Sci U S A. 1966 Aug;56(2):672–679. doi: 10.1073/pnas.56.2.672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen T. T., Heidelberger C. Quantitative studies on the malignant transformation of mouse prostate cells by carcinogenic hydrocarbons in vitro. Int J Cancer. 1969 Mar 15;4(2):166–178. doi: 10.1002/ijc.2910040207. [DOI] [PubMed] [Google Scholar]
  7. DiPaolo J. A., Donovan P., Nelson R. Quantitative studies of in vitro transformation by chemical carcinogens. J Natl Cancer Inst. 1969 May;42(5):867–874. [PubMed] [Google Scholar]
  8. DiPaolo J. A., Nelson R. L., Donovan P. J. Sarcoma-producing cell lines derived from clones transformed in vitro by benzo[a]pyrene. Science. 1969 Aug 29;165(3896):917–918. doi: 10.1126/science.165.3896.917. [DOI] [PubMed] [Google Scholar]
  9. EAGLE H., OYAMA V. I., LEVY M., FREEMAN A. E. Myo-Inositol as an essential growth factor for normal and malignant human cells in tissue culture. J Biol Chem. 1957 May;226(1):191–205. [PubMed] [Google Scholar]
  10. Freeman A. E., Kelloff G. J., Gilden R. V., Lane W. T., Swain A. P., Huebner R. J. Activation and isolation of hamster-specific C-type RNA viruses from tumors induced by cell cultures transformed by chemical carcinogens. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2386–2390. doi: 10.1073/pnas.68.10.2386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Freeman A. E., Price P. J., Bryan R. J., Gordon R. J., Gilden R. V., Kelloff G. J., Huebner R. J. Transformation of rat and hamster embryo cells by extracts of city smog. Proc Natl Acad Sci U S A. 1971 Feb;68(2):445–449. doi: 10.1073/pnas.68.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Freeman A. E., Price P. J., Igel H. J., Young J. C., Maryak J. M., Huebner R. J. Morphological transformation of rat embryo cells induced by diethylnitrosamine and murine leukemia viruses. J Natl Cancer Inst. 1970 Jan;44(1):65–78. [PubMed] [Google Scholar]
  13. Grover P. L., Sims P., Huberman E., Marquardt H., Kuroki T., Heidelberger C. In vitro transformation of rodent cells by K-region derivatives of polycyclic hydrocarbons. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1098–1101. doi: 10.1073/pnas.68.6.1098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heidelberger C., Iype P. T. Malignant transformation in vitro by carcinogenic hydrocarbons. Science. 1967 Jan 13;155(3759):214–217. [PubMed] [Google Scholar]
  15. Hsiung G. D. Activation of guinea pig C-type virus in cultured spleen cells by 5-bromo-2'-deoxyuridine. J Natl Cancer Inst. 1972 Aug;49(2):567–570. [PubMed] [Google Scholar]
  16. Huberman E., Donovan P. J., DiPaolo J. A. Mutation and transformation of cultured mammalian cells by N-acetoxy-N-2-fluorenylacetamide. J Natl Cancer Inst. 1972 Mar;48(3):837–840. [PubMed] [Google Scholar]
  17. Kamahora J., Kakunaga T. Malignant transformation of hamster embryonic cells in vitro by 4-nitroquinoline-1-oxide. Biken J. 1967 Dec;10(4):219–242. [PubMed] [Google Scholar]
  18. Kelloff G. J., Hatanaka M., Gilden R. V. Assay of C-type virus infectivity by measurement of RNA-dependent DNA polymerase activity. Virology. 1972 Apr;48(1):266–269. doi: 10.1016/0042-6822(72)90135-3. [DOI] [PubMed] [Google Scholar]
  19. Klement V., Nicolson M. O., Gilden R. V., Oroszlan S., Sarma P. S., Rongey R. W., Gardner M. B. Rat C-type virus induced in rat sarcoma cells by 5-bromodeoxyuridine. Nat New Biol. 1972 Aug 23;238(86):234–237. doi: 10.1038/newbio238234a0. [DOI] [PubMed] [Google Scholar]
  20. Klement V., Nicolson M. O., Huebner R. J. Rescue of the genome of focus forming virus from rat non-productive lines by 5'-bromodeoxyruidine. Nat New Biol. 1971 Nov 3;234(44):12–14. doi: 10.1038/newbio234012a0. [DOI] [PubMed] [Google Scholar]
  21. Kuroki T., Sato H. Transformation and neoplastic development in vitro of hamster embryonic cells by 4-nitroquinoline-1-oxide and its derivatives. J Natl Cancer Inst. 1968 Jul;41(1):53–71. [PubMed] [Google Scholar]
  22. Lowy D. R., Rowe W. P., Teich N., Hartley J. W. Murine leukemia virus: high-frequency activation in vitro by 5-iododeoxyuridine and 5-bromodeoxyuridine. Science. 1971 Oct 8;174(4005):155–156. doi: 10.1126/science.174.4005.155. [DOI] [PubMed] [Google Scholar]
  23. Namba M., Masuji H., Sato J. Carcinogenesis in tissue culture. IX. Malignant transformation of cultured rat cells treated with 4-nitroquinoline-1-oxide. Jpn J Exp Med. 1969 Jun;39(3):253–265. [PubMed] [Google Scholar]
  24. Oroszlan S., Bova D., Huebner R. J., Gilden R. V. Major group-specific protein of rat type C viruses. J Virol. 1972 Oct;10(4):746–750. doi: 10.1128/jvi.10.4.746-750.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Oroszlan S., Foreman C., Kelloff G., Gilden R. V. The group-specific antigen and other structural proteins of hamster and mouse C-type viruses. Virology. 1971 Mar;43(3):665–674. doi: 10.1016/0042-6822(71)90290-x. [DOI] [PubMed] [Google Scholar]
  26. Price P. J., Freeman A. E., Lane W. T., Huebner R. J. Morphological transformation of rat embryo cells by the combined action of 3-methylcholanthrene and Rauscher leukaemia virus. Nat New Biol. 1971 Mar 31;230(13):144–146. doi: 10.1038/newbio230144a0. [DOI] [PubMed] [Google Scholar]
  27. Rhim J. S., Creasy B., Huebner R. J. Production of altered cell foci by 3-methylcholanthrene in mouse cells infected with AKR leukemia virus. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2212–2216. doi: 10.1073/pnas.68.9.2212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rhim J. S., Vass W., Cho H. Y., Huebner R. J. Malignant transformation inducedby 7,12-dimethylbenz(a)anthracene in rat embryo cells infected with Rauscher leukemia virus. Int J Cancer. 1971 Jan 15;7(1):65–74. doi: 10.1002/ijc.2910070108. [DOI] [PubMed] [Google Scholar]
  29. Takaki R., Takii M., Ikegami T. Preliminary studies on the in vitro carcinogenesis of rat thymus cells by n-methyl-n'-nitro-n-nitrosoguanidine. Gan. 1969 Dec;60(6):661–662. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES