Skip to main content
PMC home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1995 Sep;86(9):802–810. doi: 10.1111/j.1349-7006.1995.tb03089.x

Role of Point Mutation of the K‐ras Gene in Tumorigenesis of B6C3F1 Mouse Lung Lesions Induced by Urethane

Ryoji Kawano 1, Takashi Nishisaka 1, Yukio Takeshima 1, Shuji Yonehara 1, Kouki Inai 1
PMCID: PMC5920935  PMID: 7591956

Abstract

In order to elucidate the role of point mutation of the K‐ras gene in the tumorigenetic process of lung tumors, an experimental model of lung lesions in mice induced by the administration of urethane was used. A total of 135 B6C3F1 male mice, 6 weeks old, were given urethane in the drinking water at 0, 6, 60, 600 or 1200 ppm, and were then killed after varying periods of time. The lung lesions were histologically characterized as hyperplasia, adenoma and adenocarcinoma. Point mutations in codons 12 and 61 of the K‐ras gene were detected by polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) and confirmed by using dideoxy sequencing analysis. K‐ras gene mutation was identified in 9 (23.7%) of 38 lesions classified as hyperplasia, 31 (46.3%) of 67 adenomas, and 3 (50%) of 6 adenocarcinomas. The most frequent mutation was an AT‐to‐TA transversion at the second base of codon 61 and this pattern accounted for 65% of the three mutant forms observed. These results suggest that the point mutation of K‐ras gene is involved in all stages of mouse lung tumorigenesis, i.e., activation of this gene can also influence the later stages of lung lesions.

Keywords: Urethane, Ethyl carbamate, K‐ras gene, B6C3F1 mouse, Lung tumor

Full Text

The Full Text of this article is available as a PDF (798.7 KB).

REFERENCES

  • 1. ) Sukumar , S.An experimental analysis of cancer: role of ras oncogenes in multistep carcinogenesis . Cancer Cells , 2 , 199 – 204 ( 1990. ). [PubMed] [Google Scholar]
  • 2. ) Nuzum , E. O. , Malkinson , A. M. and Beer , D. G.Specific Ki‐ras codon 61 mutations may determine the development of urethane‐induced mouse lung adenomas or adenocarcinomas . Mol. Carcinog. , 3 , 287 – 295 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Malkinson , A. M.Primary lung tumors in mice: an experimentally manipulable model of human adenocarcinoma . Cancer Res. , 52 , 2670 – 2676 ( 1992. ). [PubMed] [Google Scholar]
  • 4. ) Kumar , R. , Sukumar , S. and Barbacid , M.Activation of ras oncogenes preceding the onset of neoplasia . Science , 248 , 1101 – 1104 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Ohmori , H. , Abe , T. , Hirano , H. , Murakami , T. , Katoh , T. , Gotoh , S. , Kido , M. , Kuroiwa , A. , Nomura , T. and Higashi , K.Comparison of Ki‐ras gene mutation among simultaneously occurring multiple urethane‐induced lung tumors in individual mice . Carcinogenesis , 13 , 851 – 855 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Quintanilla , M. , Brown , K. , Ramsden , M. and Balmain , A.Carcinogen‐specific mutation and amplification of Ha‐ras during mouse skin carcinogenesis . Nature , 322 , 78 – 80 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Candrian , U. , You , M. , Goodrow , T. , Maronpot , R. R. , Reynolds , S. H. and Anderson , M. W.Activation of protooncogenes in spontaneously occurring non‐liver tumors from C57BL × C3H F1 mice . Cancer Res. , 51 , 1148 – 1153 ( 1991. ). [PubMed] [Google Scholar]
  • 8. ) You , M. , Candrian , U. , Maronpot , R. R. , Stoner , G. D. and Anderson , M. W.Activation of the K‐ras protooncogene in spontaneously occurring and chemically induced lung tumors of the strain A mouse . Proc. Natl. Acad. Sci. USA , 86 , 3070 – 3074 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. ) Goodrow , T. , Reynolds , S. , Maronpot , R. and Anderson , M.Activation of K‐ras by codon 13 mutations in C57BL/6 × C3H F1 mouse tumors induced by exposure to 1,3‐butadiene . Cancer Res. , 50 , 4818 – 4823 ( 1990. ). [PubMed] [Google Scholar]
  • 10. ) Wang , Yu. , Wang , Yi. , Stoner , G. D. and You , M.ras mutations in 2‐acetylaminofluorene‐induced lung and liver tumors from C3H/HeJ and (C3H × A/J)F1 mice . Cancer Res. , 53 , 1620 – 1624 ( 1993. ). [PubMed] [Google Scholar]
  • 11. ) Belinsky , S. A. , Devereux , T. R. , Maronpot , R. R. , Stoner , G. D. and Anderson , M. W.Relationship between the formation of promutagenic adducts and the activation of the K‐ras protooncogene in lung tumors from A/J mice treated with nitrosamines . Cancer Res. , 49 , 5305 – 5311 ( 1989. ). [PubMed] [Google Scholar]
  • 12. ) Stowers , S. J. , Glover , P. L. , Reynolds , S. H. , Boone , L. R. , Maronpot , R. R. and Anderson , M. W.Activation of the K‐ras protooncogene in lung tumors from rats and mice chronically exposed to tetranitromethane . Cancer Res. , 47 , 3212 – 3219 ( 1987. ). [PubMed] [Google Scholar]
  • 13. ) Richardson , K. K. , Rexroat , M. A. , Helvering , L. M. , Copple , D. M. and Richardson , F. C.Temporal changes in the mutant frequency and mutation spectra of the 61st codon of the H‐ras oncogene following exposure of B6C3F1 mice to N‐nitrosodiethylamine (DEN) . Carcinogenesis , 13 , 1277 – 1279 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Balmain , A. , Ramsden , M. , Bowden , G. and Smith , J.Activation of the mouse cellular Harvey‐ras gene in chemically induced benign skin papillomas . Nature , 307 , 658 – 660 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Belinsky , S. A. , Devereux , T. R. , Foley , J. F. , Maronpot , R. R. and Anderson , M. W.Role of the alveolar type II cell in the development and progression of pulmonary tumors induced by 4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanone in the A/J mouse . Cancer Res. , 52 , 3164 – 3173 ( 1992. ). [PubMed] [Google Scholar]
  • 16. ) Cerny , W. L. , Mangold , K. A. and Scarpelli , D. G.K‐ras mutation is an early event in pancreatic duct carcinogenesis in the Syrian golden hamster . Cancer Res. , 52 , 4507 – 4513 ( 1992. ). [PubMed] [Google Scholar]
  • 17. ) Tsutsumi , M. , Kondoh , S. , Noguchi , O. , Horiguchi , K. , Kobayashi , E. , Okita , S. , Ohashi , K. , Honoki , K. , Tsujiuchi , T. and Konishi , Y.K‐ras gene mutation in early ductal lesions induced in a rapid production model for pancreatic carcinomas in Syrian hamsters . Jpn. J. Cancer Res. , 84 , 1101 – 1105 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. ) Corominas , M. , Leon , J. , Kamino , H. , Cruz‐Alvarez , M. , Novick , S. C. and Pellicer , A.Oncogene involvement in tumor regression: H‐ras activation in the rabbit keratoacanthoma model . Oncogene , 6 , 645 – 651 ( 1991. ). [PubMed] [Google Scholar]
  • 19. ) Yanagisawa , A. , Ohtake , K. , Ohashi , K. , Hori , M. , Kitagawa , T. , Sugano , H. and Kato , Y.Frequent c‐Ki‐ras oncogene activation in mucous cell hyperplasias of pancreas suffering from chronic inflammation . Cancer Res. , 53 , 953 – 956 ( 1993. ). [PubMed] [Google Scholar]
  • 20. ) Strauss , B. S.The origin of point mutations in human tumor cells . Cancer Res. , 52 , 249 – 253 ( 1992. ). [PubMed] [Google Scholar]
  • 21. ) Leithauser , M. T. , Leim , A. , Stewart , B. C. , Miller , E. C. and Miller , J. A.1,N6‐Ethenoadenosine formation, mutagenicity and murine tumor induction as indicators of the generation of an electrophilic epoxide metabolite of the closely related carcinogens ethyl carbamate (urethane) and vinyl carbamate . Carcinogenesis , 11 , 463 – 473 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Dahl , G. A. , Miller , E. C. and Miller , J. A.Comparative carcinogenicities and mutagenicities of vinyl carbamate, ethyl carbamate, and ethyl n‐hydroxycarbamate . Cancer Res. , 40 , 1194 – 1203 ( 1980. ). [PubMed] [Google Scholar]
  • 23. ) Belinsky , S. A. , Foley , J. F. , White , C. M. , Anderson , M. W. and Maronpot , R. R.Dose‐response relationship between O6‐methylguanine formation in Clara cells and induction of pulmonary neoplasia in the rat by 4‐ (methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanone . Cancer Res. , 50 , 3772 – 3780 ( 1990. ). [PubMed] [Google Scholar]
  • 24. ) Stoner , G. D.Lung tumors in strain A mice as a bioassay for carcinogenicity of environmental chemicals . Exp. Lung Res. , 17 , 405 – 423 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 25. ) Saiki , R. K. , Gelfand , D. H. , Stoffel , S. , Scharf , S. J. , Higuchi , R. , Horn , G. T. , Mullis , K. B. and Erlich , H. A.Primer‐directed enzymatic amplification of DNA with a thermostable DNA polymerase . Science , 239 , 487 – 490 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Kumar , R. and Dunn , L. L.Designed diagnostic restriction fragment length polymorphisms for the detection of point mutations in ras oncogenes . Oncogene Res. , 1 , 235 – 241 ( 1989. ). [PubMed] [Google Scholar]
  • 27. ) Miller , J. A. and 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 , 48 , 1 – 15 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. ) Svensson , K.Alkylation of protein and DNA in mice treated with urethane . Carcinogenesis , 9 , 2197 – 2201 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Thaete , L. G. , Beer , D. G. and Malkinson , A. M.Genetic variation in the proliferation of murine pulmonary type II cells: basal rates and alterations following urethane treatment . Cancer Res. , 46 , 5335 – 5338 ( 1986. ). [PubMed] [Google Scholar]
  • 30. ) Malkinson , A. M. and Beer , D. S.Major effect on susceptibility to urethane‐induced pulmonary adenoma by a single gene in BALB/cBy mice . J. Natl. Cancer Inst. , 70 , 931 – 935 ( 1983. ). [PubMed] [Google Scholar]
  • 31. ) Inai , K. , Arihiro , K. , Takeshima , Y. , Yonehara , S. , Tachiyama , Y. , Khatun , N. and Nishisaka , T.Quantitative risk assessment of carcinogenicity of urethane (ethyl carbamate) on the basis of long‐term oral administration to B6C3F1 mice . Jpn. J. Cancer Res. , 82 , 380 – 385 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. ) Allen , J. W. , Stoner , G. D. , Pereira , M. A. , Backer , L. C. , Sharief , Y. , Hatch , G. G. , Campbell , J. A. , Stead , A. G. and Nesnow , S.Tumorigenesis and genotoxicity of ethyl carbamate and vinyl carbamate in rodent cells . Cancer Res. , 46 , 4911 – 4915 ( 1986. ). [PubMed] [Google Scholar]
  • 33. ) You , M. , Wang , Y. , Lineen , A. , Stoner , G. D. , You , L. , Maronpot , R. R. and Anderson , M. W.Activation of protooncogenes in mouse lung tumors . Exp. Lung Res. , 17 , 389 – 400 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Oreffo , V. C. , Lin , H. W. , Gumerlock , P. H. , Kraegel , S. A. and Witschi , H.Mutational analysis of a dominant oncogene (c‐Ki‐ras‐2) and a tumor suppressor gene (p53) in hamster lung tumorigenesis . Mol. Carcinog. , 6 , 199 – 202 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 35. ) Weinberg , R. A.Oncogenes, antioncogenes, and the molecular bases of multistep carcinogenesis . Cancer Res. , 49 , 3713 – 3721 ( 1989. ). [PubMed] [Google Scholar]
  • 36. ) Bos , J. L.ras oncogenes in human cancer: a review . Cancer Res. , 49 , 4682 – 4689 ( 1989. ). [PubMed] [Google Scholar]
  • 37. ) Land , H. , Parada , L. F. and Weinberg , R. A.Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes . Nature , 304 , 596 – 602 ( 1983. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES