Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1980 Jan;33(1):423–437. doi: 10.1128/jvi.33.1.423-437.1980

Adenovirus type 12-induced rat tumor cells of neuroepithelial origin: persistence and expression of the viral genome.

H Ibelgaufts, W Doerfler, K H Scheidtmann, W Wechsler
PMCID: PMC288558  PMID: 7365869

Abstract

Four cell lines derived from adenovirus type 12-induced rat brain tumors were studied. The polyploid cells displayed neuroepithelial characteristics and were transplantable into syngeneic rats and nude mice. In tissue culture the cells grew in monolayers and multilayers. A very high saturation density was reached, and the cells plated in agar and were easily agglutinated with low concentrations of concanavalin A. Between 2 and 11 copies of the viral genome per diploid cellular genome were detected by reassociation kinetics analysis in the different lines. The patterns of distribution of viral DNA sequences in these lines, as revealed by blot analysis, suggest colinear integration of the intact viral genome into the cellular DNA. The patterns of integration were stable after more than 15 months of prolonged tissue culture and after animal reimplantation. Integration patterns were identical in three of the tumor lines and different in another line. Viral sequences were transcribed. The extent of homology found toward adenovirus type 12 DNA in polyadenylated polysome-associated mRNA isolated from the tumor lines suggests that the early and some of the late genes of adenovirus type 12 DNA are transcribed in these tumor cells. Infectious virus was not rescuable from these lines.

Full text

PDF
424

Images in this article

Selected References

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

  1. BABLANIAN R., EGGERS H. J., TAMM I. STUDIES ON THE MECHANISM OF POLIOVIRUS-INDUCED CELL DAMAGE. I. THE RELATION BETWEEN POLIOVIRUS,-INDUCED METABOLIC AND MORPHOLOGICAL ALTERATIONS IN CULTURED CELLS. Virology. 1965 May;26:100–113. doi: 10.1016/0042-6822(65)90030-9. [DOI] [PubMed] [Google Scholar]
  2. Becker D., Kurth R., Critchley D., Friis R., Bauer H. Distinguishable transformation-defective phenotypes among temperature-sensitive mutants of Rous sarcoma virus. J Virol. 1977 Mar;21(3):1042–1055. doi: 10.1128/jvi.21.3.1042-1055.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Botchan M., Topp W., Sambrook J. The arrangement of simian virus 40 sequences in the DNA of transformed cells. Cell. 1976 Oct;9(2):269–287. doi: 10.1016/0092-8674(76)90118-5. [DOI] [PubMed] [Google Scholar]
  4. Chen L. B., Gallimore P. H., McDougall J. K. Correlation between tumor induction and the large external transformation sensitive protein on the cell surface. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3570–3574. doi: 10.1073/pnas.73.10.3570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davidson R. L., Gerald P. S. Improved techniques for the induction of mammalian cell hybridization by polyethylene glycol. Somatic Cell Genet. 1976 Mar;2(2):165–176. doi: 10.1007/BF01542629. [DOI] [PubMed] [Google Scholar]
  7. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  8. Doerfler W., Lundholm U., Rensing U., Philipson L. Intracellular forms of Adenovirus DNA. II. Isolation in dye-buoyant density gradients of a DNA-RNA complex from KB cells infected with Adenovirus type 2. J Virol. 1973 Oct;12(4):793–807. doi: 10.1128/jvi.12.4.793-807.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Druckrey H. Genotypes and phenotypes of ten inbred strains of BD-rats. Arzneimittelforschung. 1971 Aug;21(8):1274–1278. [PubMed] [Google Scholar]
  10. Fanning E., Doerfler W. Intracellular forms of adenovirus DNA. V. Viral DNA sequences in hamster cells abortively infected and transformed with human adenovirus type 12. J Virol. 1976 Nov;20(2):373–383. doi: 10.1128/jvi.20.2.373-383.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Freedman V. H., Shin S. I. Cellular tumorigenicity in nude mice: correlation with cell growth in semi-solid medium. Cell. 1974 Dec;3(4):355–359. doi: 10.1016/0092-8674(74)90050-6. [DOI] [PubMed] [Google Scholar]
  12. Freeman A. E., Calisher C. H., Price P. J., Turner H. C., Huebner R. J. Calcium sensitivity of cell cultures derived from adenovirus-induced tumors. Proc Soc Exp Biol Med. 1966 Jul;122(3):835–840. doi: 10.3181/00379727-122-31264. [DOI] [PubMed] [Google Scholar]
  13. Fujinaga K., Green M. Mechanism of viral carcinogenesis by deoxyribonucleic acid mammalian viruses. IV. Related virus-specific ribonucleic acids in tumor cells induced by "highly" oncogenic adenovirus types 12, 18, and 31. J Virol. 1967 Jun;1(3):576–582. doi: 10.1128/jvi.1.3.576-582.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gallimore P. H., McDougall J. K., Chen L. B. In vitro traits of adenovirus-transformed cell lines and their relevance to tumorigenicity in nude mice. Cell. 1977 Apr;10(4):669–678. doi: 10.1016/0092-8674(77)90100-3. [DOI] [PubMed] [Google Scholar]
  15. Gelb L. D., Kohne D. E., Martin M. A. Quantitation of Simian virus 40 sequences in African green monkey, mouse and virus-transformed cell genomes. J Mol Biol. 1971 Apr 14;57(1):129–145. doi: 10.1016/0022-2836(71)90123-9. [DOI] [PubMed] [Google Scholar]
  16. Green M. R., Chinnadurai G., Mackey J. K., Green M. A unique pattern of integrated viral genes in hamster cells transformed by highly oncogenic human adenovirus 12. Cell. 1976 Mar;7(3):419–428. doi: 10.1016/0092-8674(76)90172-0. [DOI] [PubMed] [Google Scholar]
  17. Green M. R., Mackey J. K., Green M. Multiple copies of human adenovirus 12 genomes are integrated in virus-induced hamster tumors. J Virol. 1977 Apr;22(1):238–242. doi: 10.1128/jvi.22.1.238-242.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Groneberg J., Chardonnet Y., Doerfler W. Integrated viral sequences in adenovirus type 12-transformed hamster cells. Cell. 1977 Jan;10(1):101–111. doi: 10.1016/0092-8674(77)90144-1. [DOI] [PubMed] [Google Scholar]
  19. Groneberg J., Sutter D., Soboll H., Doerfler W. Morphological revertants of adenovirus type 12-transformed hamster cells. J Gen Virol. 1978 Sep;40(3):635–645. doi: 10.1099/0022-1317-40-3-635. [DOI] [PubMed] [Google Scholar]
  20. HUEBNER R. J., ROWE W. P., LANE W. T. Oncogenic effects in hamsters of human adenovirus types 12 and 18. Proc Natl Acad Sci U S A. 1962 Dec 15;48:2051–2058. doi: 10.1073/pnas.48.12.2051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hynes R. O. Cell surface proteins and malignant transformation. Biochim Biophys Acta. 1976 Apr 30;458(1):73–107. doi: 10.1016/0304-419x(76)90015-9. [DOI] [PubMed] [Google Scholar]
  22. Ketner G., Kelly T. J., Jr Integrated simian virus 40 sequences in transformed cell DNA: analysis using restriction endonucleases. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1102–1106. doi: 10.1073/pnas.73.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee K. C., Mak S. Adenovirus type 12 DNA sequences in primary hamster tumors. J Virol. 1977 Oct;24(1):408–411. doi: 10.1128/jvi.24.1.408-411.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lindberg U., Persson T. Isolation of mRNA from KB-cells by affinity chromatography on polyuridylic acid covalently linked to Sepharose. Eur J Biochem. 1972 Dec 4;31(2):246–254. doi: 10.1111/j.1432-1033.1972.tb02527.x. [DOI] [PubMed] [Google Scholar]
  25. Lundholm U., Doerfler W. Temperature-sensitive mutants of human adenovirus type 12. Virology. 1971 Sep;45(3):827–829. doi: 10.1016/0042-6822(71)90206-6. [DOI] [PubMed] [Google Scholar]
  26. MACPHERSON I., MONTAGNIER L. AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS. Virology. 1964 Jun;23:291–294. doi: 10.1016/0042-6822(64)90301-0. [DOI] [PubMed] [Google Scholar]
  27. MCBRIDE W. D., WIENER A. IN VITRO TRANSFORMATION OF HAMSTER KIDNEY CELLS BY HUMAN ADENOVIRUS TYPE 12. Proc Soc Exp Biol Med. 1964 Apr;115:870–874. doi: 10.3181/00379727-115-29060. [DOI] [PubMed] [Google Scholar]
  28. McAllister R. M., Macpherson I. Transformation of a hamster cell line by adenovirus type 12. J Gen Virol. 1968 Jan;2(1):99–106. doi: 10.1099/0022-1317-2-1-99. [DOI] [PubMed] [Google Scholar]
  29. Mukai N., Kobayashi S. Human adenovirus-induced medulloepitheliomatous neoplasms in Sprague-Dawley rats. Am J Pathol. 1973 Dec;73(3):671–690. [PMC free article] [PubMed] [Google Scholar]
  30. Mukai N., Kobayashi S., Murao T. Cellular monstrosity in embryonic neuronal neoplasm produced by human adenovirus in rats. Acta Neuropathol. 1974;30(4):315–328. doi: 10.1007/BF00697014. [DOI] [PubMed] [Google Scholar]
  31. Mukai N., Kobayashi S. Undifferentiated intraperitoneal tumors induced by human adenovirus type 12 in hamsters. Am J Pathol. 1972 Nov;69(2):331–348. [PMC free article] [PubMed] [Google Scholar]
  32. Mukai N., Murao T. Retinal tumor induction by ocular inoculation of human adneovirus in 3-day-old rats. J Neuropathol Exp Neurol. 1975 Jan;34(1):28–35. doi: 10.1097/00005072-197501000-00003. [DOI] [PubMed] [Google Scholar]
  33. Mulder C., Sharp P. A., Delius H., Pettersson U. Specific fragmentation of DNA of adenovirus serotypes 3, 5, 7, and 12, and adeno-simian virus 40 hybrid virus Ad2+ND1 by restriction endonuclease R.EcoRI. J Virol. 1974 Jul;14(1):68–77. doi: 10.1128/jvi.14.1.68-77.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ogawa K., Hamaya K., Fujii Y., Matsuura K., Endo T. Tumor induction by adenovirus type 12 and its target cells in the central nervous system. Gan. 1969 Aug;60(4):383–392. [PubMed] [Google Scholar]
  35. Ortin J., Scheidtmann K. H., Greenberg R., Westphal M., Doerfler W. Transcription of the genome of adenovirus type 12. III. Maps of stable RNA from productively infected human cells and abortively infected and transformed hamster cells. J Virol. 1976 Nov;20(2):355–372. doi: 10.1128/jvi.20.2.355-372.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Raska K., Jr, Strohl W. A. The response of BHK21 cells to infection with type 12 adenovirus. VI. Synthesis of virus-specific RNA. Virology. 1972 Mar;47(3):734–742. doi: 10.1016/0042-6822(72)90563-6. [DOI] [PubMed] [Google Scholar]
  37. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  38. Scheidtmann K. H., Doerfler W. Transcription of the genome of adenovirus type 12. IV. Maps of stable late RNA from productively infected human cells. J Virol. 1977 Jun;22(3):585–590. doi: 10.1128/jvi.22.3.585-590.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shin S. I., Freedman V. H., Risser R., Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. doi: 10.1073/pnas.72.11.4435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  41. Sutter D., Westphal M., Doerfler W. Patterns of integration of viral DNA sequences in the genomes of adenovirus type 12-transformed hamster cells. Cell. 1978 Jul;14(3):569–585. doi: 10.1016/0092-8674(78)90243-x. [DOI] [PubMed] [Google Scholar]
  42. TRENTIN J. J., YABE Y., TAYLOR G. The quest for human cancer viruses. Science. 1962 Sep 14;137(3533):835–841. doi: 10.1126/science.137.3533.835. [DOI] [PubMed] [Google Scholar]
  43. Tjia S., Fanning E., Schick J., Doerfler W. Incomplete particles of adenovirus type 2. III. Viral and cellular DNA sequences in incomplete particles. Virology. 1977 Jan;76(1):365–379. doi: 10.1016/0042-6822(77)90309-9. [DOI] [PubMed] [Google Scholar]
  44. Trentin J. J., Van Hoosier G. L., Jr, Samper L. The oncogenicity of human adenoviruses in hamsters. Proc Soc Exp Biol Med. 1968 Mar;127(3):683–689. doi: 10.3181/00379727-127-32773. [DOI] [PubMed] [Google Scholar]
  45. Weber J. Absence of adenovirus-specific repressor in adenovirus tumour cells. J Gen Virol. 1974 Feb;22(2):259–264. doi: 10.1099/0022-1317-22-2-259. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES