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
. 1988 Jun;85(11):3673–3677. doi: 10.1073/pnas.85.11.3673

Activation of ras oncogene in aflatoxin-induced rat liver carcinogenesis.

S Sinha 1, C Webber 1, C J Marshall 1, M A Knowles 1, A Proctor 1, N C Barrass 1, G E Neal 1
PMCID: PMC280280  PMID: 3287372

Abstract

The presence of activated transforming genes was investigated in four primary aflatoxin-induced rat liver tumors in male Fischer rats, in two cell lines generated from such tumors, in an epithelial liver-derived nontransformed cell line, and in the latter cell line after transformation by aflatoxin B1 in vitro. When DNA extracted from these sources was transfected into NIH 3T3 cells, negative results were obtained from focus assays. Cotransfection of these DNA samples with a gene for resistance to G418, followed by selection for resistance to that antibiotic, and tumorigenicity testing in nude mice demonstrated DNA-mediated transfer of the neoplastic phenotype in all cases except for DNA from the nontransformed cell line. DNA extracted from these primary nude mouse tumors used in a secondary round of transfection with NIH 3T3 cells gave positive results in focus assays, which were conserved through succeeding rounds of transfection. By use of appropriate radiolabeled probes, activated ras oncogenes were detected in all samples. N-ras activation was detected in three of the primary rat liver tumors and both hepatoma cell lines. Ki-ras activation was detected in one primary rat liver tumor, and Ha-ras activation was detected in the cell line transformed in vitro with activated aflatoxin B1. The activated Ki-ras oncogene was further characterized by use of synthetic oligonucleotide probes and was shown to contain a G----A transition at the second nucleotide in codon 12.

Full text

PDF
3673

Images in this article

3675Image
on p.3675
3675Image
on p.3675
3675Image
on p.3675
3675Image
on p.3675
3676Image
on p.3676

Selected References

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

  1. Barbacid M. Oncogenes and human cancer: cause or consequence? Carcinogenesis. 1986 Jul;7(7):1037–1042. doi: 10.1093/carcin/7.7.1037. [DOI] [PubMed] [Google Scholar]
  2. Bizub D., Wood A. W., Skalka A. M. Mutagenesis of the Ha-ras oncogene in mouse skin tumors induced by polycyclic aromatic hydrocarbons. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6048–6052. doi: 10.1073/pnas.83.16.6048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bos J. L., Verlaan-de Vries M., Marshall C. J., Veeneman G. H., van Boom J. H., van der Eb A. J. A human gastric carcinoma contains a single mutated and an amplified normal allele of the Ki-ras oncogene. Nucleic Acids Res. 1986 Feb 11;14(3):1209–1217. doi: 10.1093/nar/14.3.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown R., Brady G., Mattern J., Schütz G. An alternative assay system for the detection of transforming genes. Carcinogenesis. 1984 Oct;5(10):1323–1328. doi: 10.1093/carcin/5.10.1323. [DOI] [PubMed] [Google Scholar]
  5. Bulatao-Jayme J., Almero E. M., Castro M. C., Jardeleza M. T., Salamat L. A. A case-control dietary study of primary liver cancer risk from aflatoxin exposure. Int J Epidemiol. 1982 Jun;11(2):112–119. doi: 10.1093/ije/11.2.112. [DOI] [PubMed] [Google Scholar]
  6. Butler W. H., Greenblatt M., Lijinsky W. Carcinogenesis in rats by aflatoxins B1, G1, and B2. Cancer Res. 1969 Dec;29(12):2206–2211. [PubMed] [Google Scholar]
  7. Croy R. G., Essigmann J. M., Reinhold V. N., Wogan G. N. Identification of the principal aflatoxin B1-DNA adduct formed in vivo in rat liver. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1745–1749. doi: 10.1073/pnas.75.4.1745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ellis R. W., DeFeo D., Maryak J. M., Young H. A., Shih T. Y., Chang E. H., Lowy D. R., Scolnick E. M. Dual evolutionary origin for the rat genetic sequences of Harvey murine sarcoma virus. J Virol. 1980 Nov;36(2):408–420. doi: 10.1128/jvi.36.2.408-420.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ellis R. W., Defeo D., Shih T. Y., Gonda M. A., Young H. A., Tsuchida N., Lowy D. R., Scolnick E. M. The p21 src genes of Harvey and Kirsten sarcoma viruses originate from divergent members of a family of normal vertebrate genes. Nature. 1981 Aug 6;292(5823):506–511. doi: 10.1038/292506a0. [DOI] [PubMed] [Google Scholar]
  10. Eva A., Aaronson S. A. Frequent activation of c-kis as a transforming gene in fibrosarcomas induced by methylcholanthrene. Science. 1983 May 27;220(4600):955–956. doi: 10.1126/science.6302839. [DOI] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Garner R. C., Martin C. N., Smith J. R., Coles B. F., Tolson M. R. Comparison of aflatoxin B1 and aflatoxin G1 binding to cellular macromolecules in vitro, in vivo and after peracid oxidation; characterisation of the major nucleic acid adducts. Chem Biol Interact. 1979 Jun;26(1):57–73. doi: 10.1016/0009-2797(79)90093-0. [DOI] [PubMed] [Google Scholar]
  13. Garte S. J., Hood A. T., Hochwalt A. E., D'Eustachio P., Snyder C. A., Segal A., Albert R. E. Carcinogen specificity in the activation of transforming genes by direct-acting alkylating agents. Carcinogenesis. 1985 Dec;6(12):1709–1712. doi: 10.1093/carcin/6.12.1709. [DOI] [PubMed] [Google Scholar]
  14. Gu J. R., Tian P. K., Wan D. F., Wang X., Li H. N., Pan Z. M., Huang L. H., Li X. Z., Jiang H. Q. Identification of human N-ras as the common oncogene in NIH 3T3 cells transformed with DNAs from human primary hepatic cancer and hepatoma 7402 line. Sci Sin B. 1986 Feb;29(2):173–180. [PubMed] [Google Scholar]
  15. Harris C. C., Sun T. Multifactoral etiology of human liver cancer. Carcinogenesis. 1984 Jun;5(6):697–701. doi: 10.1093/carcin/5.6.697. [DOI] [PubMed] [Google Scholar]
  16. Manson M. M., Legg R. F., Watson J. V., Green J. A., Neal G. E. An examination of the relative resistances to aflatoxin B1 and susceptibilities to gamma-glutamyl p-phenylene diamine mustard of gamma-glutamyl transferase negative and positive cell lines. Carcinogenesis. 1981;2(7):661–670. doi: 10.1093/carcin/2.7.661. [DOI] [PubMed] [Google Scholar]
  17. Martin C. N., Garner R. C. Aflatoxin B -oxide generated by chemical or enzymic oxidation of aflatoxin B1 causes guanine substitution in nucleic acids. Nature. 1977 Jun 30;267(5614):863–865. doi: 10.1038/267863a0. [DOI] [PubMed] [Google Scholar]
  18. McCann J., Ames B. N. Detection of carcinogens as mutagens in the Salmonella/microsome test: assay of 300 chemicals: discussion. Proc Natl Acad Sci U S A. 1976 Mar;73(3):950–954. doi: 10.1073/pnas.73.3.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McMahon G., Davis E., Wogan G. N. Characterization of c-Ki-ras oncogene alleles by direct sequencing of enzymatically amplified DNA from carcinogen-induced tumors. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4974–4978. doi: 10.1073/pnas.84.14.4974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McMahon G., Hanson L., Lee J. J., Wogan G. N. Identification of an activated c-Ki-ras oncogene in rat liver tumors induced by aflatoxin B1. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9418–9422. doi: 10.1073/pnas.83.24.9418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Miller E. C. Some current perspectives on chemical carcinogenesis in humans and experimental animals: Presidential Address. Cancer Res. 1978 Jun;38(6):1479–1496. [PubMed] [Google Scholar]
  22. Ochiya T., Fujiyama A., Fukushige S., Hatada I., Matsubara K. Molecular cloning of an oncogene from a human hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1986 Jul;83(14):4993–4997. doi: 10.1073/pnas.83.14.4993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Padua R. A., Barrass N. C., Currie G. A. Activation of N-ras in a human melanoma cell line. Mol Cell Biol. 1985 Mar;5(3):582–585. doi: 10.1128/mcb.5.3.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Peers F. G., Linsell C. A. Dietary aflatoxins and liver cancer--a population based study in Kenya. Br J Cancer. 1973 Jun;27(6):473–484. doi: 10.1038/bjc.1973.60. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Quintanilla M., Brown K., Ramsden M., Balmain A. Carcinogen-specific mutation and amplification of Ha-ras during mouse skin carcinogenesis. Nature. 1986 Jul 3;322(6074):78–80. doi: 10.1038/322078a0. [DOI] [PubMed] [Google Scholar]
  26. Reynolds S. H., Stowers S. J., Maronpot R. R., Anderson M. W., Aaronson S. A. Detection and identification of activated oncogenes in spontaneously occurring benign and malignant hepatocellular tumors of the B6C3F1 mouse. Proc Natl Acad Sci U S A. 1986 Jan;83(1):33–37. doi: 10.1073/pnas.83.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sinha S., Hockin L. J., Neal G. E. A system for transformation of rat liver cells in vitro by acute treatment with aflatoxin. Br J Cancer. 1987 Jun;55(6):595–598. doi: 10.1038/bjc.1987.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sinha S., Marshall C. J., Neal G. E. gamma-Glutamyltranspeptidase and the ras-induced transformation of a rat liver cell line. Cancer Res. 1986 Mar;46(3):1440–1445. [PubMed] [Google Scholar]
  29. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  30. Sukumar S., Notario V., Martin-Zanca D., Barbacid M. Induction of mammary carcinomas in rats by nitroso-methylurea involves malignant activation of H-ras-1 locus by single point mutations. Nature. 1983 Dec 15;306(5944):658–661. doi: 10.1038/306658a0. [DOI] [PubMed] [Google Scholar]
  31. Swenson D. H., Lin J. K., Miller E. C., Miller J. A. Aflatoxin B1-2,3-oxide as a probable intermediate in the covalent binding of aflatoxins B1 and B2 to rat liver DNA and ribosomal RNA in vivo. Cancer Res. 1977 Jan;37(1):172–181. [PubMed] [Google Scholar]
  32. Wang T. C., Cerutti P. A. Effect of formation and removal of aflatoxin B1:DNA adducts in 10T 1/2 mouse embryo fibroblasts on cell viability. Cancer Res. 1980 Aug;40(8 Pt 1):2904–2909. [PubMed] [Google Scholar]
  33. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wiseman R. W., Stowers S. J., Miller E. C., Anderson M. W., Miller J. A. Activating mutations of the c-Ha-ras protooncogene in chemically induced hepatomas of the male B6C3 F1 mouse. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5825–5829. doi: 10.1073/pnas.83.16.5825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Witney F. R., Furano A. V. Highly repeated DNA families in the rat. J Biol Chem. 1984 Aug 25;259(16):10481–10492. [PubMed] [Google Scholar]
  36. Wogan G. N., Newberne P. M. Dose-response characteristics of aflatoxin B1 carcinogenesis in the rat. Cancer Res. 1967 Dec;27(12):2370–2376. [PubMed] [Google Scholar]
  37. Zarbl H., Sukumar S., Arthur A. V., Martin-Zanca D., Barbacid M. Direct mutagenesis of Ha-ras-1 oncogenes by N-nitroso-N-methylurea during initiation of mammary carcinogenesis in rats. 1985 May 30-Jun 5Nature. 315(6018):382–385. doi: 10.1038/315382a0. [DOI] [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