Summary
In order to assess the possibility that activatedras-associated hepatic carcinomas might be much rarer in rats than mice because of the more frequent or rapid occurrence of powerful carcinogenic event(s) other thanras point mutations in the former animals, precancerous lesions and hepatocellular carcinomas induced by a weak hepatocarcinogenN-methyl-N-nitrosourea (MNU) in the rat liver were analyzed for the presence or absence ofras point mutations. MNU was chosen because it is well known that MNU-induced rat mammary carcinomas contain activated H-ras at very high frequency. Male Fisher rats were treated with a single dose of MNU after partial hepatectomy, and then administered dietary phenobarbital or repeated s.c. injections of carbon tetrachloride as promoting procedures. Analyses by oligonucleotide hybridization, MnlI-restriction-fragment-length polymorphism and NIH3T3 cell transfection assays revealed neither H-ras point mutations nor transforming ability of the DNA from 36 MNU-induced rat hepatic neoplasms. The results were in agreement with previous results for rat hepatocellular carcinomas induced by other potent liver carcinogens and did not support our hypothesis that the frequency of findingras activation might be dependent on the strength of the carcinogen.
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Abbreviations
- MNU:
-
N-methyl-N-nitrosourea
- HCC:
-
hepatocellular carcinoma
References
Balmain A, Brown K (1988) Oncogene activation in chemical carcinogenesis. Adv Cancer Res 51:147–182
Barbacid M (1987) Ras genes. Annu Rev Biochem 56:779–827
Buchmann A, Mahr J, Bauer-Hofmann R, Schwarz M (1989) Mutations at codon 61 of the Ha-ras Proto-oncogene in precancerous liver lesions of the B6C3F1 mouse. Mol Carcinog 2:121–125
Butler WH, Newberne PM (eds) (1975) Mouse hepatic neoplasia. Elsevier, Amsterdam
Farber E (1984) Cellular biochemistry of the stepwise development of cancer with chemicals: GHA, Clowes Memorial Lecture. Cancer Res 44:5463–5474
Funato T, Yokota J, Sakamoto H, Kameya T, Fukushima S, Ho N, Terada M, Sugimura T (1987) Activation of N-ras gene in a rat hepatocellular carcinoma induced by dibutylnitrosamine and butylate hydroxytoluene. Jpn J Cancer Res (Gann) 78:689–694
Harper JR, Roop DR, Yuspa SH (1986) Transfection of the EJras Ha gene into keratinocytes derived from carcinogen-induced mouse papillomas causes malignant progression. Mol Cell Biol 6:3144–3149
Hsu SM, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled PAP procedures. J Histochem Cytochem 29:577–580
Ishikawa F, Takaku F, Nagao M, Ochiai M, Hayashi K, Takayama S, Sugimura T (1985) Activated oncogenes in a rat hepatocellular carcinoma induced by 2-amino-3-methylimidazo(4,5-f)-quinoline. Jpn J Cancer Res 76:425–428
Kitagawa T, Sugano H (1978) Enhancing effect of phenobarbital on the development of enzyme-altered islands and hepatocellular carcinomas initiated by 3′-methyl-4′-dimethylaminoazobenzene or diethylnitrosamine. Gann 69:679–689
Kitagawa T, Hirakawa T, Ishikawa T, Nemoto N, Takayama S (1980) Induction of hepatocellular carcinoma in rat liver by initial treatment with benzo(a)pyrene after partial hepatectomy and promotion by phenobarbital. Toxicol Lett 6:167–171
Knudson AG (1985) Hereditary cancer, oncogenes, and anti-oncogenes. Cancer Res 45:1437–1443
Kumar R, Barbacid M (1988) Oncogene detection at the single cell level. Oncogene 3:647–651
Maekawa A, Onodera H, Kanno J, Furuta K, Nagaoka T, Todate A, Matsushima Y, Oh-hara T, Kawazoe Y (1988) Carcinogenicity and organ specificity ofN-trimethylsilylmethyl-N-nitrosourea-(TMS-MNU),N-neopentyl-N-nitrosourea-(neoPNU), andN-methyl-N-nitrosourea (MNU) in rats. J Cancer Res Clin Oncol 114:473–476
McCormic DL, Adamowski CB, Fiks A, Moon RC (1981) Lifetime dose-response relationship for mammary tumor induction by a single administration of N-methyl-N-nitrosourea. Cancer Res 41:1690–1694
McMahon G, Hanson L, Lee J, Wogan GN (1986) Identification of an activated c-Ki-ras oncogene in rat liver tumors induced by aflatoxin B1. Proc Natl Acad Sci USA 83:9148–9422
Moore MA, Kitagawa T (1986) Hepatocarcinogenesis in the rat: the effect of promoters and carcinogens in vivo and in vitro. Int Rev Cytol 101:125–173
Ramsden M, Cole G, Smith J, Balmain A (1985) Differential methylation of the c-H-ras gene in normal mouse cells and during skin tumour progression. EMBO J 4:1449–1454
Reynolds SH, Stowers SJ, Moronpot RR, Anderson MW (1986) Differential and identification of activated oncogenes in spontaneously occuring benign and malignant hepatocellular tumors of the B6C3F1 mouse. Proc Natl Acad Sci USA 83:33–37
Saiki RK, Scharf S, Faloona F, Millis KB, Horn GT, Erlich A, Arnheim N (1985) Enzymatic amplification ofγ-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230:1350–1354
Sato K (1989) GlutathioneS-transferase and hepatocarcinogenesis. Adv Cancer Res 52:205–255
Sinha S, Webber C, Marshall CJ, Knowles MA, Proctor A, Barrass NC, Neal GE (1988) Activation ofras oncogene in aflatoxin-induced rat liver carcinogenesis. Proc Natl Acad Sci USA 85:3673–3677
Slamon DJ, Cline MJ (1984) Expression of cellular oncogenes during embryonic and fetal development of the mouse. Proc Natl Acad Sci USA 81:7141–7145
Stanislawski BM, Uriel J, Graber P (1967) Association of embryonic antigens with experimentally induced hepatic lesions in the rats. Cancer Res 27:1900–1997
Stowers SJ, Wiseman RW, Ward JM, Miller EC, Miller JA, Anderson MW, Eva A (1988) Detection of activated proto-oncogenes inN-nitrosodiethylamine-induced liver tumors: a comparison between B6C3F1 mice and Fischer 344 rats. Carcinogenesis 9:271–276
Sukumar S, Notario V, Martin-Zanca D, Barbacid M (1983) Induction of mammary carcinomas in rats by nitroso-methylurea involves malignant activation of H-ras-1 locus by single point mutations. Nature 306:658–661
Tsuda H, Fukushima S, Imaida K, Kurata Y, Ito N (1983) Organ-specific promoting effects of phenobarbital and saccharin in induction of thyroid, liver, and urinary tumors in rats after initiation withN-nitrosomethylurea. Cancer Res 43:3292–3296
Verlaan-de VM, Bogaard ME, Van den EH, Van Boom JH, Van der Eb AJ, Bos JL (1986) A dot-blot screening procedure for mutated ras oncogenes using synthetic oligodeoxynucleotides. Gene 50:313–320
Watatani M, Perantoni AO, Reed CD, Enomoto T, Wenk ML, Rice JM (1989) Infrequent activation of K-ras, H-ras, and other oncogenes in hepatocellular neoplasms initiated by methyl(acetoxymethyl)nitrosamine, a methylating agent, and promoted by phenobarbital in F344 rats. Cancer Res 49:1103–1109
Wiseman RW, Stowers SJ, Miller EC, Anderson MW, Miller JA (1986) Activating mutations of c-H-ras protooncogene in chemically induced hepatomas of the male B6C3F1 mouse. Proc Natl Acad Sci USA 83:5825–5829
Wiseman RW, Stewart BC, Grenier D, Miller EC, Miller JA (1987) Characterization of c-H-ras proto-oncogene in chemically induced hepatomas of the B6C3F1 mouse. Proc Am Assoc Cancer Res 28, abstr 585
Zarbl H, Sukumar S, Arthur AV, Martin-Zanca D, Barbacid M (1985) Direct mutagenesis of H-ras-1 oncogenes byN-nitroso-N-methylurea during initiation of mammary carcinogenesis in rats. Nature 315:382–385
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Li, H., Lee, G.H., Cui, L. et al. Absence of H-ras point mutation at codon 12 inN-methyl-N-nitrosourea-induced hepatocellular neoplasms in the rat. J Cancer Res Clin Oncol 116, 331–335 (1990). https://doi.org/10.1007/BF01612914
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DOI: https://doi.org/10.1007/BF01612914