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. 1997 Feb;88(2):129–136. doi: 10.1111/j.1349-7006.1997.tb00357.x

Demonstration of ras and p53 Gene Mutations in Carcinomas in the Forestomach and Intestine and Soft Tissue Sarcomas Induced by N‐Methyl‐N‐nitrosourea in the Rat

Kazuyuki Matsumoto 1, Teruhiko Iwase 1, Iwao Hirono 1, Yoshihisa Nishida 2, Yoshio Iwahori 3, Takaaki Hori 3, Makoto Asamoto 3, Nobuo Takasuka 3, Dae Joong Kim 3, Toshikazu Ushijima 4, Minako Nagao 4, Hiroyuki Tsuda 3,✉
PMCID: PMC5921361  PMID: 9119740

Abstract

The presence of ras family and p53 gene mutations in rat forestomach, intestine and liver tumors and soft tissue sarcomas induced by Nmethyl‐N‐nitrosourea (MNU) was examined using polymerase chain reaction‐single strand conformation polymorphism (PCR‐SSCP) followed by direct sequencing analysis. In the forestomach squamous cell carcinomas (SCC), Ha‐ros and p53 mutations were detected in 2 (40%) and 4 (80%) of 5 cases, respectively. The figures for Ki‐ras and p53 gene mutations in adenocarcinomas of the large and small intestines were 3 (18.8%) and 5 (31.3%) of 16 cases. Soft tissue sarcomas in different sites were found to have mutations of Ki‐ras in 7 (23.3%)and of p53 in 9 (30%) of 30 cases. One forestomach SCC and 2 soft tissue sarcomas had double p53 mutations in different exons. Single cases of forestomach SCC and intestinal adenocarcinoma had mutations in both Ki‐ras and p53 genes. No mutations were found in counterpart benign tumors or hepatocellular adenomas. The p53 mutation spectrum revealed preferential clustering within exon 8 for the forestomach SCCs, and exons 5 and 8 for the intestinal adenocarcinomas, whereas the distribution was evenly spread through exons 5 to 8 in soft tissue sarcomas. All the detected ras or p53 mutations were G:C to A:T transitions. These results indicate firstly that specific Ki‐ras, Ha‐ras and p53 gene mutations in MNU‐induced lesions are related to particular alkylation sites (G:C to A:T transitions) and secondly, although not essential, Ki‐ras, Ha‐ras or p53 gene mutations may be involved in the progression stage of forestomach, intestine and soft tissue neoplasms induced by MNU.

Keywords: pS3, ras, Tumors, MNU, Rat

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REFERENCES

  • 1. ) Fearon , E. R. and Vogelstein , B.A genetic model for colorectal tumorigenesis . Cell , 61 , 759 – 767 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Forrester , K. , Almoguera , C , Han , K. , Grizzle , W. E. and Perucho , M.Detection of high incidence of K‐rasoncogenes during human colon tumorigenesis . Nature , 327 , 298 – 303 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Bos , J. L. , Fearon , E. R. , Hamilton , S. R. , Verlaan‐de , V. M. , van Boom , J. H. , van der Eb , A. J. and Vogelstein , B.Prevalence of ras gene mutations in human colorectal cancers . Nature , 327 , 293 – 297 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Llor , S. , Jacoby , R. P. , Teng , B.‐B. , Davidson , N. O. , Sitrin , M. D. and Brasitus , T. A.K‐ras mutations in 1,2‐dimethylhydrazine‐induced colonic tumors: effects of supplemental dietary calcium and vitamin D deficiency . Cancer Res. , 51 , 4305 – 4309 ( 1991. ). [PubMed] [Google Scholar]
  • 5. ) Stopera , S. A. , Murphy , L. C. and Bird , R. P.Evidence of a ras gene mutation in azoxymethane‐induced colonic aberrant crypts in Sprague Dawley rats. Earliest recognizable precursor lesion of experimental colon cancer . Carcinogenesis , 13 , 2081 – 2085 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Sukumar , S. , Armstrong , B. , Bruyntjes , J. P. , Leav , I. and Bosland , M. C.Frequent activation of the Ki ras oncogene at codon 12 in N‐methyl‐N‐nitrosourea‐induced rat prostate adenocarcinomas and neurogenic sarcomas . Mol Carcinog. , 4 , 362 – 368 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Jacoby , R. F. , Alexander , R. J. , Raicht , R. F. and Brasitus , T. A.K‐ras oncogene mutations in rat colon tumors induced by N‐methyl‐N‐nitrosourea . Carcinogenesis , 13 , 45 – 49 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Baker , S. J. , Preisinger , A. C. , Jessup , J. M. , Paraskeva , C. , Markowitz , S. , Willson , J. K. , Hamilton , S. and Vogelstein , B.p53 gene mutations occur in combination with 17p allelic deletions as late events in colorectal tumorigenesis . Cancer Res. , 50 , 7717 – 7722 ( 1990. ). [PubMed] [Google Scholar]
  • 9. ) Greenblatt , M. S. , Bennet , W. P. , Hollstein , M. and Harris , C. C.Mutation in the p53 tumor suppressor gene: clue to cancer etiology and molecular pathogenesis . Cancer Res. , 54 , 4855 – 4878 ( 1994. ). [PubMed] [Google Scholar]
  • 10. ) Shivapurkar , N. , Belinsky , A. , Wolf , D. C. , Tang , Z. and Alabaster , O.Absence of p53 gene mutations in rat colon carcinomas induced by azoxymethane . Cancer Lett. , 96 , 63 – 70 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Toguchida , J. , Yamaguchi , T. , Ritchie , B. , Beauchamp , R. L. , Dayton , S. H. , Herrera , G. E. , Yamamuro , T. , Kotoura , Y. , Sasaki , M. S. , Little , J. B. , Weichselbaum , R. R. , Ishizaki , K. and Yandell , D. W.Mutation spectrum of the p53 gene in bone and soft tissue sarcomas . Cancer Res. , 52 , 6194 – 6199 ( 1992. ). [PubMed] [Google Scholar]
  • 12. ) Ohgaki , H , Hard , G. C , Hirota , N. , Maekawa , A. , Takahashi , M. and Kleihues , P.Selective mutation of codons 204 and 213 of the p53 gene in rat tumors induced by alkylating N‐nitroso compounds . Cancer Res. , 52 , 2995 – 2998 ( 1992. ). [PubMed] [Google Scholar]
  • 13. ) Weghorst , C. M. , Dragnev , K. H. , Buzard , G. S. , Thorne , K. L. , Vandeborne , G. F. , Vincent , K. A. and Rice , J. M.Low incidence of point mutations detected in the p53 tumor suppressor gene from chemically induced rat renal mesenchymal tumors . Cancer Res. , 54 , 215 – 219 ( 1994. ). [PubMed] [Google Scholar]
  • 14. ) Harvey , M. , McArthur , M. J. , MontgomeryJr. , C. A. , Butel , J. S. , Bradley , A. , and Donehower , A.Spontaneous and carcinogen‐induced tumorigenesis in p53‐deficient mice . Nat Genet. , 5 , 225 – 229 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Bursting , S. D. , Perking , S. N. and Phang , J. M.Calorie restriction delays spontaneous tumorigenesis in p53‐knockout transgenic mice . Proc. Natl. Acad. Sci. USA , 91 , 7036 – 7040 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. ) Pudie , C. A. , Harrison , D. J. , Peter , A. , Bobbie , L. , White , S. , Howie , S. E. M. , Salter , D. M. , Bird , C. C. , Whyllie , A. H. , Hooper , M. L. , and Clarke , A. R.Tumor incidence, spectrum and ploidy in mice with a large deletion in the p53 gene . Oncogene , 9 , 603 – 609 ( 1994. ). [PubMed] [Google Scholar]
  • 17. ) Makino , H. , Ishizaka , Y. , Tsujimoto , A. , Nakamura , T. , Onda , M. , Sugimura , T. and Nagao , M.Rat p53 gene mutations in primary Zymbal gland tumors induced by 2‐amino‐3‐methylimidazo[4,5‐f]quinoline, a food mutagen . Proc. Natl. Acad. Sci. USA , 89 , 4850 – 4854 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. ) Masui , T. , Mann , A. M. , Macatee , T. L. , Okamura , T. , Garland , E. M. , Smith , R. A. and Cohen , S. M.Absence of ras oncogene activation in rat urinary bladder carcinomas induced by N‐methyl‐N‐nitrosourea or N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine . Carcinogenesis , 13 , 2281 – 2285 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) Hulla , J. E.The rat genome contains a p53 pseudogene: detection of a processed pseudogene using PCR . PCR Methods Appl , 1 , 251 – 254 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Zarbl , H. , Sukumar , S. , Arthur , A. V. , Martin , Z. D. and Barbacid , M.Direct mutagenesis of Ha‐ras‐1 oncogenes by N‐nitroso‐N‐methylurea during initiation of mammary carcinogenesis in rats . Nature , 315 , 382 – 385 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Sukumar , S. , Notario , V. , Martin , Z. D. and Barbacid , M.Induction of mammary carcinomas in rats by nitrosomethylurea involves malignant activation of H‐ras‐1 locus by single point mutations . Nature , 306 , 658 – 661 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Lozano , J. C. , Nakazawa , H. , Cros , M. P. , Cabral , R. and Yamasaki , H.G—Amutations inp53 and Ha‐ras genes in esophageal papillomas induced by N‐nitrosomethylbenzyl‐amine in two strains of rats . Mol. Carcinog. , 9 , 33 – 39 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Sakai , H. and Ogawa , K.Mutational activation of H‐ras and K‐ ras genes is absent in N‐nitroso‐N‐methylurea‐induced liver tumors in rats . Jpn, J. Cancer Res. , 81 , 437 – 439 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. ) Li , H. , Lee , G. H. , Cui , L. X. , Liu , J. , Nomura , K. , Ohtake , K. and Kitagawa , T.Absence of H‐ ras point mutation at codon 12 in N‐methyl‐N‐nitrosourea‐induced hepatoceUular neoplasms in the rat . J. Cancer Res. Clin. Oncol , 116 , 331 – 335 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 25. ) Matsumoto , K. , Tsuda , H. , Iwase , T. , Ito , M. , Nishida , Y. , Oyama , F. , Titani , K. , Ushijima , T. , Nagao , M. and Hirono , I.Absence of ras family point mutations at codon 12, 13 and 61 in N‐ethyl‐N‐hydroxyethylnitrosamine‐ or N‐nitrosomorpholine‐induced renal cell tumors in rats . Jpn. J. Cancer Res. 83 , 933 – 936 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. ) Smith , M. L. , Yeleswarapu , L. , Scalamogna , P. , Locker , J. and Lombardi , B.p53 mutations in hepatoceUular carcinomas induced by a choline‐devoid diet in male Fischer 344 rats . Carcinogenesis , 14 , 503 – 510 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Recio , L. , Sisk , S. , Pluta , L. , Bermudez , E. , Gross , E. A. , Chen , Z. , Morgan , K. and Walker , C.p53 mutations in formaldehyde‐induced nasal squamous cell carcinomas in rats . Cancer Res. , 52 , 6113 – 6116 ( 1992. ). [PubMed] [Google Scholar]
  • 28. ) Vancutsem , P. M. , Lazarus , P. and Williams , G.Frequent and specific mutations of the rat p53 gene in hepatocarcinogenesis induced by tamoxifen . Cancer Res. , 54 , 3864 – 3867 ( 1994. ). [PubMed] [Google Scholar]
  • 29. ) Tokusashi , Y. , Fukuda , I. and Ogawa , K.Absence of p53 mutations and various frequencies of Ki‐ras exon 1 mutations in rat hepatic tumors induced by different carcinogens . Mol. Carcinog. , 10 , 45 – 51 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Masui , T. , Don , I. , Takada , N. , Ogawa , K. , Shirai , T. and Fukushima , S.p53 mutations in early neoplastic lesions of the urinary bladder in rats treated with N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine . Carcinogenesis , 15 , 2379 – 2381 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Asamoto , M. , Mann , A. M. , Macatee , T. L. and Cohen , S. M.Mutations and expression of the p53 gene in rat bladder carcinomas and cell lines . Mol. Carcinog. , 9 , 236 – 244 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Hollstein , M. , Sidransky , D. , Vogelstein , B. and Harris , C. C.p53 mutations in human cancers [Review] . Science , 253 , 49 – 53 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Shibata , M. A. , Shirai , T. , Ogawa , K. , Takahashi , S. , Wild , C. P. , Montesano , R. , Tsuda , H. and Ito , N.DNA methylation adduct formation and H‐ ras gene mutations in progression of N‐butyl‐N‐(4‐hydroxybutyl)nitrosamineinduced bladder tumors caused by a single exposure to N‐methyl‐N‐nitrosourea . Carcinogenesis , 15 , 2965 – 2968 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Singer , B. and Kusmierek , J. T.Chemical mutagenesis . Annu. Rev. Siochem. , 51 , 655 – 693 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 35. ) Richardson , K. K. , Richardson , F. C. , Crosby , R. M. , Swenberg , J. A. and Skopek , T. R.DNA base changes and alkylation following in vivo exposure of Escherichia coli to N‐methyl‐N‐nitrosourea or N‐ethyl‐N‐nitrosourea . Proc. Natl. Acad. Set. USA , 84 , 344 – 348 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36. ) Vogelstein , B. , Fearon , E. R. , Hamilton , S. R. , Kern , S. E. , Preisinger , A. C. , Leppert , M. , Nakamura , Y. , White , R. , Smits , A. M. and Bos , J. L.Genetic alterations during colorectal‐tumor development . N. Engl. J. Med. , 319 , 525 – 532 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 37. ) Balmain , A. and Brown , K.Oncogene activation in chemical carcinogenesis . Adv. Cancer Res. , 51 , 147 – 182 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 38. ) Ito , N. , Hasegawa , R. , Imaida , K. , Hirose , M. , Asamoto , M. and Shirai , T.Concepts in multistage carcinogenesis . Crit. Rev. Oncol/Hematol 21 , 105 – 133 ( 1995. ). [DOI] [PubMed] [Google Scholar]

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