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. 1986 Aug;83(16):5825–5829. doi: 10.1073/pnas.83.16.5825

Activating mutations of the c-Ha-ras protooncogene in chemically induced hepatomas of the male B6C3 F1 mouse.

R W Wiseman, S J Stowers, E C Miller, M W Anderson, J A Miller
PMCID: PMC386388  PMID: 3016723

Abstract

Activated c-Ha-ras protooncogenes have recently been identified in the DNA of some spontaneous hepatic tumors found in 2-year-old B6C3 F1 mice. Activation of c-Ha-ras has now been demonstrated in DNA from well-differentiated hepatomas initiated by a single dose of carcinogen given to male B6C3 F1 mice at 12 days of age. DNA from each of 25 hepatomas, induced by N-hydroxy-2-acetylaminofluorene, vinyl carbamate, or 1'-hydroxy-2',3'-dehydroestragole, containing transforming activity in the NIH 3T3 transfection assay. Southern analysis of NIH 3T3 cells transformed by DNA from 24 of these hepatomas revealed amplified and/or rear-ranged restriction fragments homologous to a Ha-ras probe. The other tumor contained an activated Ki-ras gene. Immunoprecipitation and NaDodSO4/PAGE analysis of p21 ras proteins in NIH 3T3 transformants derived from a majority of the hepatomas suggested that the activating mutations were localized in the 61st codon of the c-Ha-ras gene. Creation of a new Xba I restriction site by an AT----TA transversion at the second position of codon 61 was detected in DNA from primary tumors and NIH 3T3 cells transformed by DNA from 6 of 7 vinyl carbamate- and 5 of 10 1'-hydroxy-2',3'-dehydroestragole-induced hepatomas. Selective oligonucleotide hybridization demonstrated a CG----AT transversion at the first position of the 61st codon in NIH 3T3 transformants derived from 7 of 7 N-hydroxy-2-acetylaminofluorene-induced hepatomas. By the same criterion, an AT----GC transition at the second position of codon 61 was the activating mutation in 1 of 7 vinyl carbamate- and 5 of 10 1'-hydroxy-2',3'-dehydroestragole-induced tumors. Thus, c-Ha-ras activation is apparently an early event in B6C3 F1 mouse hepatocarcinogenesis that results directly from reaction of ultimate chemical carcinogens with this gene in vivo.

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Selected References

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

  1. Andersen P. R., Tronick S. R., Aaronson S. A. Structural organization and biological activity of molecular clones of the integrated genome of a BALB/c mouse sarcoma virus. J Virol. 1981 Nov;40(2):431–439. doi: 10.1128/jvi.40.2.431-439.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Balmain A., Ramsden M., Bowden G. T., Smith J. Activation of the mouse cellular Harvey-ras gene in chemically induced benign skin papillomas. Nature. 1984 Feb 16;307(5952):658–660. doi: 10.1038/307658a0. [DOI] [PubMed] [Google Scholar]
  3. Balmain A. Transforming ras oncogenes and multistage carcinogenesis. Br J Cancer. 1985 Jan;51(1):1–7. doi: 10.1038/bjc.1985.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barbin A., Besson F., Perrard M. H., Béréziat J. C., Kaldor J., Michel G., Bartsch H. Induction of specific base-pair substitutions in E. coli trpA mutants by chloroethylene oxide, a carcinogenic vinyl chloride metabolite. Mutat Res. 1985 Nov-Dec;152(2-3):147–156. doi: 10.1016/0027-5107(85)90056-9. [DOI] [PubMed] [Google Scholar]
  5. Bos J. L., Toksoz D., Marshall C. J., Verlaan-de Vries M., Veeneman G. H., van der Eb A. J., van Boom J. H., Janssen J. W., Steenvoorden A. C. Amino-acid substitutions at codon 13 of the N-ras oncogene in human acute myeloid leukaemia. 1985 Jun 27-Jul 3Nature. 315(6022):726–730. doi: 10.1038/315726a0. [DOI] [PubMed] [Google Scholar]
  6. Bos J. L., Verlaan-de Vries M., Jansen A. M., Veeneman G. H., van Boom J. H., van der Eb A. J. Three different mutations in codon 61 of the human N-ras gene detected by synthetic oligonucleotide hybridization. Nucleic Acids Res. 1984 Dec 11;12(23):9155–9163. doi: 10.1093/nar/12.23.9155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Der C. J., Finkel T., Cooper G. M. Biological and biochemical properties of human rasH genes mutated at codon 61. Cell. 1986 Jan 17;44(1):167–176. doi: 10.1016/0092-8674(86)90495-2. [DOI] [PubMed] [Google Scholar]
  8. Dhar R., Ellis R. W., Shih T. Y., Oroszlan S., Shapiro B., Maizel J., Lowy D., Scolnick E. Nucleotide sequence of the p21 transforming protein of Harvey murine sarcoma virus. Science. 1982 Sep 3;217(4563):934–936. doi: 10.1126/science.6287572. [DOI] [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. Fasano O., Aldrich T., Tamanoi F., Taparowsky E., Furth M., Wigler M. Analysis of the transforming potential of the human H-ras gene by random mutagenesis. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4008–4012. doi: 10.1073/pnas.81.13.4008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fennell T. R., Wiseman R. W., Miller J. A., Miller E. C. Major role of hepatic sulfotransferase activity in the metabolic activation, DNA adduct formation, and carcinogenicity of 1'-hydroxy-2',3'-dehydroestragole in infant male C57BL/6J x C3H/HeJ F1 mice. Cancer Res. 1985 Nov;45(11 Pt 1):5310–5320. [PubMed] [Google Scholar]
  12. Fox T. R., Watanabe P. G. Detection of a cellular oncogene in spontaneous liver tumors of B6C3F1 mice. Science. 1985 May 3;228(4699):596–597. doi: 10.1126/science.3983645. [DOI] [PubMed] [Google Scholar]
  13. Furth M. E., Davis L. J., Fleurdelys B., Scolnick E. M. Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family. J Virol. 1982 Jul;43(1):294–304. doi: 10.1128/jvi.43.1.294-304.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  15. Guerrero I., Villasante A., Corces V., Pellicer A. Loss of the normal N-ras allele in a mouse thymic lymphoma induced by a chemical carcinogen. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7810–7814. doi: 10.1073/pnas.82.23.7810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kidd V. J., Wallace R. B., Itakura K., Woo S. L. alpha 1-antitrypsin deficiency detection by direct analysis of the mutation in the gene. Nature. 1983 Jul 21;304(5923):230–234. doi: 10.1038/304230a0. [DOI] [PubMed] [Google Scholar]
  17. Lai C. C., Miller J. A., Miller E. C., Liem A. N-sulfoöxy-2-aminofluorene is the major ultimate electrophilic and carcinogenic metabolite of N-hydroxy-2-acetylaminofluorene in the livers of infant male C57BL/6J x C3H/HeJ F1 (B6C3F1) mice. Carcinogenesis. 1985 Jul;6(7):1037–1045. doi: 10.1093/carcin/6.7.1037. [DOI] [PubMed] [Google Scholar]
  18. Laib R. J., Gwinner L. M., Bolt H. M. DNA alkylation by vinyl chloride metabolites: etheno derivatives or 7-alkylation of guanine? Chem Biol Interact. 1981 Oct;37(1-2):219–231. doi: 10.1016/0009-2797(81)90179-4. [DOI] [PubMed] [Google Scholar]
  19. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  20. Miller E. C., Miller J. A., Boberg E. W., Delclos K. B., Lai C. C., Fennell T. R., Wiseman R. W., Liem A. Sulfuric acid esters as ultimate electrophilic and carcinogenic metabolites of some alkenylbenzenes and aromatic amines in mouse liver. Carcinog Compr Surv. 1985;10:93–107. [PubMed] [Google Scholar]
  21. Miller E. C., Miller J. A. Searches for ultimate chemical carcinogens and their reactions with cellular macromolecules. Cancer. 1981 May 15;47(10):2327–2345. doi: 10.1002/1097-0142(19810515)47:10<2327::aid-cncr2820471003>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  22. Miller J. A., Miller E. C. The metabolic activation and nucleic acid adducts of naturally-occurring carcinogens: recent results with ethyl carbamate and the spice flavors safrole and estragole. Br J Cancer. 1983 Jul;48(1):1–15. doi: 10.1038/bjc.1983.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ramsden M., Cole G., Smith J., Balmain A. Differential methylation of the c-H-ras gene in normal mouse cells and during skin tumour progression. EMBO J. 1985 Jun;4(6):1449–1454. doi: 10.1002/j.1460-2075.1985.tb03801.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reddy E. P., Lipman D., Andersen P. R., Tronick S. R., Aaronson S. A. Nucleotide sequence analysis of the BALB/c murine sarcoma virus transforming gene. J Virol. 1985 Mar;53(3):984–987. doi: 10.1128/jvi.53.3.984-987.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reddy E. P., Reynolds R. K., Santos E., Barbacid M. A point mutation is responsible for the acquisition of transforming properties by the T24 human bladder carcinoma oncogene. Nature. 1982 Nov 11;300(5888):149–152. doi: 10.1038/300149a0. [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. Seeburg P. H., Colby W. W., Capon D. J., Goeddel D. V., Levinson A. D. Biological properties of human c-Ha-ras1 genes mutated at codon 12. Nature. 1984 Nov 1;312(5989):71–75. doi: 10.1038/312071a0. [DOI] [PubMed] [Google Scholar]
  28. Singer B., Abbott L. G., Spengler S. J. Assessment of mutagenic efficiency of two carcinogen-modified nucleosides, 1,N6-ethenodeoxyadenosine and O4-methyldeoxythymidine, using polymerases of varying fidelity. Carcinogenesis. 1984 Sep;5(9):1165–1171. doi: 10.1093/carcin/5.9.1165. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Srivastava S. K., Yuasa Y., Reynolds S. H., Aaronson S. A. Effects of two major activating lesions on the structure and conformation of human ras oncogene products. Proc Natl Acad Sci U S A. 1985 Jan;82(1):38–42. doi: 10.1073/pnas.82.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Varmus H. E. The molecular genetics of cellular oncogenes. Annu Rev Genet. 1984;18:553–612. doi: 10.1146/annurev.ge.18.120184.003005. [DOI] [PubMed] [Google Scholar]
  33. Vousden K. H., Bos J. L., Marshall C. J., Phillips D. H. Mutations activating human c-Ha-ras1 protooncogene (HRAS1) induced by chemical carcinogens and depurination. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1222–1226. doi: 10.1073/pnas.83.5.1222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yuasa Y., Srivastava S. K., Dunn C. Y., Rhim J. S., Reddy E. P., Aaronson S. A. Acquisition of transforming properties by alternative point mutations within c-bas/has human proto-oncogene. Nature. 1983 Jun 30;303(5920):775–779. doi: 10.1038/303775a0. [DOI] [PubMed] [Google Scholar]
  35. 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]

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