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. 1991 Feb;87(2):624–630. doi: 10.1172/JCI115039

Mutations in the K-ras oncogene induced by 1,2-dimethylhydrazine in preneoplastic and neoplastic rat colonic mucosa.

R F Jacoby 1, X Llor 1, B B Teng 1, N O Davidson 1, T A Brasitus 1
PMCID: PMC296352  PMID: 1991846

Abstract

These experiments were conducted to determine whether point mutations activating K-ras or H-ras oncogenes, induced by the procarcinogen 1,2-dimethylhydrazine (DMH), were detectable in preneoplastic or neoplastic rat colonic mucosa. Rats were injected weekly with diluent or DMH at 20 mg/kg body wt for 5, 10, 15, or 25 wk, killed, and their colons dissected. DNA was extracted from diluent-injected control animals, histologically normal colonic mucosa from carcinogen-treated animals, and from carcinomas. Ras mutations were characterized by differential hybridization using allele-specific oligonucleotide probes to polymerase chain reaction--amplified DNA, and confirmed by DNA sequencing. While no H-ras mutations were detectable in any group, K-ras (G to A) mutations were found in 66% of DMH-induced colon carcinomas. These mutations were at the second nucleotide of codons 12 or 13 or the first nucleotide of codon 59 of the K-ras gene. The same type of K-ras mutations were observed in premalignant colonic mucosa from 2 out of 11 rats as early as 15 wk after beginning carcinogen injections when no dysplasia, adenomas, or carcinomas were histologically evident, suggesting that ras mutation may be an early event in colon carcinogenesis.

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Selected References

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

  1. Ahnen D. J. Are animal models of colon cancer relevant to human disease. Dig Dis Sci. 1985 Dec;30(12 Suppl):103S–106S. doi: 10.1007/BF01296987. [DOI] [PubMed] [Google Scholar]
  2. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  3. Barkla D. H., Tutton P. J. Ultrastructure of 1,2-dimethylhydrazine-induced adenocarcinomas in rat colon. J Natl Cancer Inst. 1978 Nov;61(5):1291–1299. doi: 10.1093/jnci/61.5.1291. [DOI] [PubMed] [Google Scholar]
  4. Bos J. L., Fearon E. R., Hamilton S. R., Verlaan-de Vries M., van Boom J. H., van der Eb A. J., Vogelstein B. Prevalence of ras gene mutations in human colorectal cancers. 1987 May 28-Jun 3Nature. 327(6120):293–297. doi: 10.1038/327293a0. [DOI] [PubMed] [Google Scholar]
  5. Bos J. L. ras oncogenes in human cancer: a review. Cancer Res. 1989 Sep 1;49(17):4682–4689. [PubMed] [Google Scholar]
  6. Burmer G. C., Loeb L. A. Mutations in the KRAS2 oncogene during progressive stages of human colon carcinoma. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2403–2407. doi: 10.1073/pnas.86.7.2403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caignard A., Kitagawa Y., Sato S., Nagao M. Activated K-ras in tumorigenic and non-tumorigenic cell variants from a rat colon adenocarcinoma, induced by dimethylhydrazine. Jpn J Cancer Res. 1988 Feb;79(2):244–249. doi: 10.1111/j.1349-7006.1988.tb01583.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Delattre O., Olschwang S., Law D. J., Melot T., Remvikos Y., Salmon R. J., Sastre X., Validire P., Feinberg A. P., Thomas G. Multiple genetic alterations in distal and proximal colorectal cancer. Lancet. 1989 Aug 12;2(8659):353–356. doi: 10.1016/s0140-6736(89)90537-0. [DOI] [PubMed] [Google Scholar]
  9. Forrester K., Almoguera C., Han K., Grizzle W. E., Perucho M. Detection of high incidence of K-ras oncogenes during human colon tumorigenesis. 1987 May 28-Jun 3Nature. 327(6120):298–303. doi: 10.1038/327298a0. [DOI] [PubMed] [Google Scholar]
  10. Freeman H. J., Kim Y., Kim Y. S. Glycoprotein metabolism in normal proximal and distal rat colon and changes associated with 1,2-dimethylhydrazine-induced colonic neoplasia. Cancer Res. 1978 Oct;38(10):3385–3390. [PubMed] [Google Scholar]
  11. George D. L., Scott A. F., Trusko S., Glick B., Ford E., Dorney D. J. Structure and expression of amplified cKi-ras gene sequences in Y1 mouse adrenal tumor cells. EMBO J. 1985 May;4(5):1199–1203. doi: 10.1002/j.1460-2075.1985.tb03760.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LaMont J. T., O'Gorman T. A. Experimental colon cancer. Gastroenterology. 1978 Dec;75(6):1157–1169. [PubMed] [Google Scholar]
  13. Lynch P. M., Lynch H. T., Harris R. E. Hereditary proximal colonic cancer. Dis Colon Rectum. 1977 Nov-Dec;20(8):661–668. doi: 10.1007/BF02586685. [DOI] [PubMed] [Google Scholar]
  14. Maskens A. P., Dujardin-Loits R. M. Experimental adenomas and carcinomas of the large intestine behave as distinct entities: most carcinomas arise de novo in flat mucosa. Cancer. 1981 Jan 1;47(1):81–89. doi: 10.1002/1097-0142(19810101)47:1<81::aid-cncr2820470115>3.0.co;2-t. [DOI] [PubMed] [Google Scholar]
  15. Mitra G., Pauly G. T., Kumar R., Pei G. K., Hughes S. H., Moschel R. C., Barbacid M. Molecular analysis of O6-substituted guanine-induced mutagenesis of ras oncogenes. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8650–8654. doi: 10.1073/pnas.86.22.8650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pegg A. E. Methylation of the O6 position of guanine in DNA is the most likely initiating event in carcinogenesis by methylating agents. Cancer Invest. 1984;2(3):223–231. doi: 10.3109/07357908409104376. [DOI] [PubMed] [Google Scholar]
  17. Perucho M., Goldfarb M., Shimizu K., Lama C., Fogh J., Wigler M. Human-tumor-derived cell lines contain common and different transforming genes. Cell. 1981 Dec;27(3 Pt 2):467–476. doi: 10.1016/0092-8674(81)90388-3. [DOI] [PubMed] [Google Scholar]
  18. Ruta M., Wolford R., Dhar R., Defeo-Jones D., Ellis R. W., Scolnick E. M. Nucleotide sequence of the two rat cellular rasH genes. Mol Cell Biol. 1986 May;6(5):1706–1710. doi: 10.1128/mcb.6.5.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Tabor S., Richardson C. C. Effect of manganese ions on the incorporation of dideoxynucleotides by bacteriophage T7 DNA polymerase and Escherichia coli DNA polymerase I. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4076–4080. doi: 10.1073/pnas.86.11.4076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tabor S., Richardson C. C. Selective inactivation of the exonuclease activity of bacteriophage T7 DNA polymerase by in vitro mutagenesis. J Biol Chem. 1989 Apr 15;264(11):6447–6458. [PubMed] [Google Scholar]
  24. Tahira T., Hayashi K., Ochiai M., Tsuchida N., Nagao M., Sugimura T. Structure of the c-Ki-ras gene in a rat fibrosarcoma induced by 1,8-dinitropyrene. Mol Cell Biol. 1986 Apr;6(4):1349–1351. doi: 10.1128/mcb.6.4.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Thurnherr N., Deschner E. E., Stonehill E. H., Lipkin M. Induction of adenocarcinomas of the colon in mice by weekly injections of 1,2-dimethylhydrazine. Cancer Res. 1973 May;33(5):940–945. [PubMed] [Google Scholar]
  26. Topal M. D. DNA repair, oncogenes and carcinogenesis. Carcinogenesis. 1988 May;9(5):691–696. doi: 10.1093/carcin/9.5.691. [DOI] [PubMed] [Google Scholar]
  27. Vogelstein B., Fearon E. R., Hamilton S. R., Kern S. E., Preisinger A. C., Leppert M., Nakamura Y., White R., Smits A. M., Bos J. L. Genetic alterations during colorectal-tumor development. N Engl J Med. 1988 Sep 1;319(9):525–532. doi: 10.1056/NEJM198809013190901. [DOI] [PubMed] [Google Scholar]
  28. Ward J. M. Morphogenesis of chemically induced neoplasms of the colon and small intestine in rats. Lab Invest. 1974 Apr;30(4):505–513. [PubMed] [Google Scholar]
  29. Weinberg R. A. Oncogenes, antioncogenes, and the molecular bases of multistep carcinogenesis. Cancer Res. 1989 Jul 15;49(14):3713–3721. [PubMed] [Google Scholar]
  30. Wood W. I., Gitschier J., Lasky L. A., Lawn R. M. Base composition-independent hybridization in tetramethylammonium chloride: a method for oligonucleotide screening of highly complex gene libraries. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1585–1588. doi: 10.1073/pnas.82.6.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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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