Skip to main content
SpringerLink
Log in

Detection of chemical carcinogens by assay of unscheduled DNA synthesis in rat tracheal epithelium in short-term organ culture

  • Original Papers
  • Experimental Oncology
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Summary

A short-term organ culture of rat tracheal epithelium was used to detect the ability of 53 chemicals to induce UDS. In this system all direct-acting compounds (ultimate or proximate carcinogens) tested induced UDS. Of 24 compounds requiring metabolism (procarcinogens), nine induced UDS, viz., 4NQO, AF-2, BP, DMN, DEN, and NP. Urethane, AAF, and 2,7-AAF induced very slight UDS. 3-Methyl-4NQO for which carcinogenicity data is incomplete was positive in our system. Among the cancer chemotherapeutic agents tested only mitomycin C induced UDS. MC and DMBA, which are known to induce cancer of respiratory organs in experimental animals, and DAB, aflatoxin B1 and Trp-P-1, which are strong carcinogens in the liver, did not induce UDS within 2 h. With the longer exposure (24 h), these carcinogens also failed to elicit UDS. All the carcinogens that induce UDS showed clear dose-dependent effects. No non-carcinogens tested induced UDS. These results suggested that this system should be useful for screening environmental chemicals suspected of damaging DNA of the respiratory organ on the basis of organotropic effects for UDS induction in cultured rat tracheal epithelium.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

UDS:

unscheduled DNA synthesis

4NQO:

4-nitroquinoline 1-oxide

Af-2:

2-(2-furyl)-3(5-nitro-2-furyl)acrylamide

BP:

benzo(a)pyrene

DMN:

dimethylnitrosamine

DEN:

diethylnitrosamine

NP:

N-nitrosopiperidine

AAF:

2-acetylaminofluorene

2,7-AAF:

2,7-bisacetylaminofluorene

MC:

3-methylcholanthrene

DMBA:

7,12-dimethylbenz(a)anthracene

DAB:

3′-methyl-4-dimethylaminoazobenzene

Trp-P-1:

3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indole

MMS:

methylmethane sulfonate

EMS:

ethylmethane sulfonate

DPN:

di-N-propylnitrosamine

DBN:

di-N-butylnitrosamine

MNU:

N-methyl-N-nitrosourea

BNU:

N-butyl-N-nitrosourea

MNNG:

N-methyl-N′-nitro-N-nitrosoguanidine

TPA:

12-O-tetradecanoyl-phorbol-13-acetate

[methyl-3H]dThd:

[methyl-3H]thymidine

ENU:

N-ethyl-N-nitrosourea

ENNG:

N-ethyl-N′-nitro-N-nitrosoguanidine

MAMacetate:

methylazoxymethanol acetate

N-OH AAF:

N-hydroxy-2-acetylaminofluorene

Trp-P-2:

3-amino-1-methyl-5H-pyride(4,3-b)indole

Glu-P-1:

2-amino-6-methyldipyride(1,2-a:3′,2′-d)imidazole

Glu-P-2:

2-amino-dipyride(1,2-a:3′,2′-d)imidazole

4AQO:

4-aminoquinoline 1-oxide

References

  • Autrup H, Essigman JM, Croy RG, Trump BF, Wogan GN, Harris CC (1979) Metabolism of alfatoxin B1 and identification of the major aflatoxin B1-DNA adducts formed in cultured human bronchus and colon. Cancer Res 39:694–698

    Google Scholar 

  • Autrup H, Wefald FC, Jeffery AM, Tate H, Schwartz RD, Trump BF, Harris CC (1980) Metabolism of benzo(a)pyrene by cultured tracheobronchial tissues from mice, rats, hamsters, bovines, and humans. Int J Cancer 25:293–300

    Google Scholar 

  • Brouns RE, Poot M, Vrind R, Hoek-Kon, Henderson P (1979) Measuremen of DNA-excision repair in suspension of freshly isolated rat hepatocytes after exposure to some carcinogenic compounds; Its possible use in carcinogenicity screening. Mutation Res 64:425–432

    Google Scholar 

  • Brown HS, Jeffery AM, Weinstein IB (1979) Formation of DNA adducts in 10T1/2 mouse embryo fibroblasts incubated with benzo(a)pyrene or dihydrodiol oxide derivatives. Cancer Res 39:1673–1677

    Google Scholar 

  • Ciarrochi G, Linn S (1978) A cell free assay measuring DNA synthesis in human fibroblasts. Proc Natl Acad Sci USA 75:1887–1891

    Google Scholar 

  • Clayson DB, Garner RC (1976) Carcinogenic aromatic amines and related compounds. In: Searle CE (ed) Chemical carcinogens. American Chemical Society, Washington, pp 434–435

    Google Scholar 

  • Cohen GM, Marchok AC, Nettesheim P, Steele VE, Nelson F, Huang S, Selkirk JK (1979) Comparative metabolism of benzo(a)pyrene in organ and cell cultures derived from rat tracheas. Cancer Res 39:1980–1984

    Google Scholar 

  • Cohen GM, Moore BP (1976) Metabolism of [3H]benzo(a)pyrene by different portions of the respiratory tract. Biochem Pharmacol 25:1623–1629

    Google Scholar 

  • Eisenbrand G, Spiegelhalder B, Janzowski C, Kann J, Preussmann R (1978) Volatile and non-volatile N-nitroso compounds in food and other environmental media. In: Walker EA, Griciute L, Castegnaro M, Lyle RE, Davis W (eds) Environmental aspects of N-nitroso compounds. International Agency for Research on Cancer, Lyon, France, pp 311–324

    Google Scholar 

  • Feldman HJ, Remsen J, Shinohara K, Cerutti P (1978) Excisability and persistence of benzo(a)pyrene DNA adducts in epithelioid human lung cells. Nature 274:796–798

    Google Scholar 

  • Freeman HJ, San RHC (1980) Use of unscheduled DNA synthesis in freshly isolated human intestinal mucosal cells for carcinogen detection. Cancer Res 40:3155–3157

    Google Scholar 

  • Harris CC (1977) Respiratory carcinogenesis. In: Straus MJ (ed) Lung cancer. Clinical diagnosis and treatment. Grune and Stratton, New York, pp 1–17

    Google Scholar 

  • Harris CC, Autrup H, Connor R, Barrett LA, McDowell EM, Trump BF (1976a) Interindividual variation in binding of benzo(a)pyrene to DNA in cultured human bronchi. Science 194:1067–1069

    Google Scholar 

  • Harris CC, Autrup H, Stoner GD, McDowell EB, Trump BF, Shafer P (1977a) Metabolism of acyclic and cyclic N-nitrosamines in cultured human bronchi. J Natl Cancer Inst 59:1401–1406

    Google Scholar 

  • Harris CC, Autrup H, Stoner GD, Trump BF (1978) Carcinogenesis studies in human respiratory epithelium. An experimental model system. In: Harris CC (ed) Pathogenesis and therapy of lung cancer, Marcel Dekker New York, pp 559–607

    Google Scholar 

  • Harris CC, Autrup H, Stoner GD, Yang SK Leutz JC, Gelboin HV, Selkirk JK, Connor RJ, Barett LA, Jones RT, McDowell E, Trump BF (1977b) Metabolism of benzo(a)pyrene and 7,12-dimethyl-benz(a)anthracene in cultured human bronchus and pancreatic duct. Cancer Res 37:3349–3355

    Google Scholar 

  • Harris CC, Frank AL, Haaften C, Kaufman DG, Connor R, Jackson F, Barrett LA, McDowell EM, Trump BF (1976b) Binding of [3H]-benzo(a)pyrene to DNA in cultured human bronchus. Cancer Res 36:1011–1018

    Google Scholar 

  • Harris CC, Genta VM, Frank AL, Kaufman DG, Barrett LA, McDowell EM, Trump BF (1974) Carcinogenic polynuclear hydrocarbons bind to macromolecules in cultured human bronchi. Nature 252:68–69

    Google Scholar 

  • Higginson J (1977) The role of the pathologist in environmental medicine and public health. Am J Pathol 86:459–484

    Google Scholar 

  • Higginson J, Muir CS (1973) Epidemiology. In: Holland JF, Rei E (eds) Cancer medicine, Lea and Febiger, Philadelphia, pp 241–307

    Google Scholar 

  • Hollstein M, McCann J, Angelosanto FA, Nicholas WW (1979) Short-term tests for carcinogens and mutagens. Mutation Res 65:133–226

    Google Scholar 

  • IARC Monographs on the evaluation of the carcinogenic risk of chemicals to man (1971–1976) Vols. 1–12. International Agency for Research on Cancer, Lyon, France

  • Ishikawa T, Takayama S, Ide F (1980) Autoradiographic demonstration of DNA repair synthesis in rat tracheal epithelium treated with chemical carcinogens in vitro. Cancer Res 40:2898–2903

    Google Scholar 

  • Ishikawa T, Takayama S, Kitagawa T, Kawachi T, Kinebuchi M, Matsukara N, Uchida E, Sugimura T (1979) In vivo experiments on tryptophan pyrolysis products. In: Miller EC, Miller JA, Hirono I, Sugimura T, Takayama S (eds) Naturally occurring carcinogens-mutagens and modulators of carcinogenesis. Japan Scientific Societies Press, Tokyo, pp 159–167

    Google Scholar 

  • Laishes BA, Koropatonick DJ, Stich HF (1975) Organ-specific DNA damage induced in mice by the organotropic carcinogens 4-nitroquinoline 1-oxide and dimethylnitrosamine. Proc Soc Exp Biol Med 149:978–982

    Google Scholar 

  • Laishes, BA, Stich HF (1973) Repair synthesis and sedimentation analysis of DNA of human cells exposed to dimethylnitrosamine and activated dimethylnitrosamine. Biochem Biophys Res Commun 52:827–833

    Google Scholar 

  • Lake RS, Kropko ML, McLachlan S, Pezzutti MR, Schoemarker RH, Igel HJ (1980) Chemical carcinogen induction of DNA-repair synthesis in human peripheral blood monocytes. Mutation Res 74:357–377

    Google Scholar 

  • Lake RS, Kropko ML, Pezzutti MR, Schoemarker RH, Igel HJ (1978) Chemical induction of unscheduled DNA synthesis in human skin epithelial cell cultures. Cancer Res 38:2091–2098

    Google Scholar 

  • Lieberman MW, Poirier MC (1973) Deoxyribonucleotide incorporation during DNA repair of carcinogen-induced damage in human diploid fibroblasts. Cancer Res 33:2097–2103

    Google Scholar 

  • Magee PN, Montesano R, Preussman R (1976) N-nitroso compounds and related carcinogens. In: Searle CE (ed) Chemical carcinogens. American Chemical Society, Washington, pp 491–625

    Google Scholar 

  • Marquart H, Philips FS, Sternberg SS (1976) Tumorigenicity in vivo and induction of malignant transformation and mutagenesis in cell cultures by adriamycin and daunomycin. Cancer Res 36:2065–2069

    Google Scholar 

  • Martin CN, McDermid AC, Garner RC (1978) Testing of known carcinogens and non-carcinogens for their ability to induce unscheduled DNA synthesis in Hela cells. Cancer Res 38:2621–2627

    Google Scholar 

  • McCann J, Ames BN (1976) Detection of carcinogens as mutagens in the Salmonella/microsome test: Assay of 300 chemicals. Discussion. Proc Natl Acad Sci USA 73:950–954

    Google Scholar 

  • McCann J, Choi E, Yamasaki E, Ames BN (1976) Detecting of carcinogens as mutagens in the Salmonella/microsome test: Assay of 300 chemicals. Proc Natl Acad Sci USA 72:5135–5139

    Google Scholar 

  • Michalopoulas G, Sattler GL, O'Conner L, Pitot HC (1978) Unscheduled DNA synthesis induced by procarcinogens in suspensions and primary cultures of hepatocytes on collagen membrane. Cancer Res 38:1866–1871

    Google Scholar 

  • Moore BP, Cohen GM (1978) Metabolism of benzo(a)pyrene and its major metabolites to ethyl acetatesoluble and water-soluble metabolites by cultured rodent trachea. Cancer Res 38:3066–3075

    Google Scholar 

  • Nettesheim P, Griesemer RA (1978) Experimental models for studies of respiratory tract carcinogenesis. In: Harris CC (ed) Pathogenesis and therapy of lung cancer. Marcel Dekker, New York, pp 75–188

    Google Scholar 

  • Nettesheim P, Marchok A, Terazaghi M (1978) In vivo and in vitro studies of the evaluation of epithelial neoplasia. In: Gelboin HV, Ts'O POP (eds) Polycyclic hydrocarbons and cancer, Vol 2. Academic Press, New York, pp 307–329

    Google Scholar 

  • Painter RB (1977) Rapid test to detect agents that damage human DNA. Nature 265:650–651

    Google Scholar 

  • Peleg L, Raz E, Ben-Ishai R (1976) Changing capacity for DNA excision repair in mouse embryonic cells in vitro. Exp Cell Res 104:301–307

    Google Scholar 

  • Petzold GL, Swenberg JA (1978) Detection of DNA damage induced in vivo following exposure of rats to carcinogens. Cancer Res 38:1589–1594

    Google Scholar 

  • Rasmussen PE, Painter RB (1964) Evidence for repair of ultraviolet damaged deoxyribonucleic acid in cultured mammalian cells. Nature 203:1360–1362

    Google Scholar 

  • San RHC, Stich HF (1975) DNA repair synthesis of cultured human cells as a rapid bioassay for chemical carcinogens. Int J Cancer 16:284–291

    Google Scholar 

  • Sirica AE, Hwang CG, Sattler GL, Pitot HC (1980) Use of primary cultures of adult rat hepatocytes on collagen gel-nylon mesh to evaluate carcinogen-induced unscheduled DNA synthesis. Cancer Res 40:3259–3267

    Google Scholar 

  • Stich HF, Kieser D (1974) Use of DNA repair synthesis in detecting organotropic actions of chemical carcinogens. Proc Soc Exp Biol Med 145:1339–1342

    Google Scholar 

  • Stich HF, Kieser D, Laishes BA, San RHC, Warren P (1975) DNA repair of human cells as a relevant, rapid, and economic assay for environmental carcinogens. Gann Monograph on Cancer Res 17:3–15

    Google Scholar 

  • Stich HF, San RHC, Kawazoe Y (1971) DNA repair synthesis in mammalian cells exposed to a series of oncogenic and non-oncogenic derivatives of 4-nitroquinoline 1-oxide. Nature 229:416–419

    Google Scholar 

  • Stich HF, San RHC, Lam PPS, Koropatonick DJ, Lo LW, Laishes BA (1976) DNA fragmentation and DNA repair as an in vitro and in vivo assay for chemical procarcinogens, carcinogens and carcinogenic nitrosation products. In: Montesano R, Bartsch H, Tomatis L (eds) Screening tests in chemical carcinogenesis. International Agency for Research on Cancer, Lyon, France, pp 639–655

    Google Scholar 

  • Stoltz DR, Poirier LA, Irving CC, Stich HF, Weisburger JH, Grice HC (1974) Evaluation of short-term tests for carcinogenicity. Tox Appl Pharmacol 29:157–180

    Google Scholar 

  • Sugimura T, Fujimura S (1967) Tumor production in glandular stomach of rat by N-methyl-N′-nitro-N-nitrosoguanidine. Nature 216:943–944

    Google Scholar 

  • Svoboda D, Reddy J, Harris C (1970) Invasive Tumors induced in rats with actinomycin D. Cancer Res 30:2271–2279

    Google Scholar 

  • Tannenbaum SR (1979) Endogenous formation on nitrite and N-nitroso compounds. In: Miller EC, Miller JA, Hirono I, Sugimura T, Takayama S (eds) Naturally occurring carcinogens-mutagens and modulators of carcinogenesis. Japan Scientific Societies Press, Tokyo, pp 211–220

    Google Scholar 

  • Williams GM (1977) Detection of chemical carcinogens by unscheduled DNA synthesis in rat liver primary cell cultures. Cancer Res 37:1845–1851

    Google Scholar 

  • Yager JD, Miller JA (1978) DNA repair in primary cultures of rat hepatocytes. Cancer Res 38:4385–4394

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dept. of Experimental Pathology, Cancer Institute, 1-37-1, Kami-Ikebukuro Toshima-ku, 170, Tokyo, Japan

    F. Ide, T. Ishikawa & S. Takayama

Authors
  1. F. Ide
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Ishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Takayama
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by a grant-in-aid for cancer research from the Ministry of Education, Science, and Culture, and grants from the Society for Promotion of Cancer Research, and the Japan Tobacco and Salt Public Corporation of Japan

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ide, F., Ishikawa, T. & Takayama, S. Detection of chemical carcinogens by assay of unscheduled DNA synthesis in rat tracheal epithelium in short-term organ culture. J Cancer Res Clin Oncol 102, 115–126 (1981). https://doi.org/10.1007/BF00410663

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00410663

Key words

Search

Navigation