Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Oct;79(19):6028–6032. doi: 10.1073/pnas.79.19.6028

alpha-Difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase, inhibits tumor promoter-induced polyamine accumulation and carcinogenesis in mouse skin.

C E Weeks, A L Herrmann, F R Nelson, T J Slaga
PMCID: PMC347045  PMID: 6821130

Abstract

The role of ornithine decarboxylase (OrnDCase, EC 4.1.1.17) and of the polyamines [putrescine (Put), spermidine (Spd), and spermine (Spm)] in mouse skin tumor promotion was investigated by the use of alpha-difluoromethylornithine (CHF2-Orn), an enzyme-activated irreversible inhibitor of OrnDCase. 12-O-Tetradecanoylphorbol 13-acetate (TPA), mezerein, and ethyl phenylpropiolate (EPP) were employed as complete, stage II specific, and nonpromoting agents, respectively. TPA and mezerein, but not EPP, provided for a dose-dependent increase in tissue Put accumulation. The Put level in papillomas developed by TPA (2 micrograms) treatment was approximately equal to 15-fold higher than that of the surrounding skin tissue; Spd accumulation was 2- to 3-fold greater in the papillomas. Put administered (intraperitoneally) with TPA greatly enhanced papilloma yield. CHF2-Orn, given orally or intraperitoneally, abolished the TPA-induced OrnDCase activity and Put accumulation in mouse epidermis. The reduction of polyamine accumulation by CHF2-Orn was directly proportional to reduction of tumor size. CHF2-Orn administered in a two-stage (TPA-mezerein) promotion protocol [Slaga, T. J., Fischer, S. M., Nelson, K. G. & Gleason, G. L. (1980) Proc. Natl. Acad. Sci. USA 77, 3659-3663; Slaga, T. J., Klein-Szanto, A. J. P., Fischer, S. M., Weeks, C. E., Nelson, K. & Major, S. (1980) Proc. Natl. Acad. Sci. USA 77, 2251-2254] reduced tumor size, inhibited by 65-70% the number of papillomas per mouse, and decreased by 40% the percentage of mice with tumors when given with the stage II agent mezerein. CHF2-Orn provided considerably less effect on tumorigenesis when administered with the TPA portion of the protocol, and CHF2-Orn did not inhibit the induction of dark basal keratinocytes by TPA. Based on our results with CHF2-Orn, we suggest that regulation of polyamine biosynthesis, particularly Put, is a critical factor in stage II promotion.

Full text

PDF
6028

Images in this article

6031Image
on p.6031

Selected References

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

  1. Bacchi C. J., Nathan H. C., Hutner S. H., McCann P. P., Sjoerdsma A. Polyamine metabolism: a potential therapeutic target in trypanosomes. Science. 1980 Oct 17;210(4467):332–334. doi: 10.1126/science.6775372. [DOI] [PubMed] [Google Scholar]
  2. Bartholeyns J., Koch-Weser J. Effects of alpha-difluoromethylornithine alone and combined with adriamycin or vindesine on L1210 leukemia in mice, EMT6 solid tumors in mice, and solid tumors induced by injection of hepatoma tissue culture cells in rats. Cancer Res. 1981 Dec;41(12 Pt 1):5158–5161. [PubMed] [Google Scholar]
  3. Emanuelsson H., Heby O. Inhibition of putrescine synthesis blocks development of the polychete Ophryotrocha labronica at gastrulation. Proc Natl Acad Sci U S A. 1978 Feb;75(2):1039–1042. doi: 10.1073/pnas.75.2.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fozard J. R., Part M. L., Prakash N. J., Grove J., Schechter P. J., Sjoerdsma A., Koch-Weser J. L-Ornithine decarboxylase:an essential role in early mammalian embryogenesis. Science. 1980 May 2;208(4443):505–508. doi: 10.1126/science.6768132. [DOI] [PubMed] [Google Scholar]
  5. Gazitt Y., Friend C. Polyamine biosynthesis enzymes in the induction and inhibition of differentiation in Friend erythroleukemia cells. Cancer Res. 1980 May;40(5):1727–1732. [PubMed] [Google Scholar]
  6. Huberman E., Weeks C., Herrmann A., Callaham M., Slaga T. Alterations in polyamine levels induced by phorbol diesters and other agents that promote differentiation in human promyelocytic leukemia cells. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1062–1066. doi: 10.1073/pnas.78.2.1062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jänne J., Pösö H., Raina A. Polyamines in rapid growth and cancer. Biochim Biophys Acta. 1978 Apr 6;473(3-4):241–293. doi: 10.1016/0304-419x(78)90015-x. [DOI] [PubMed] [Google Scholar]
  8. Klein-Szanto A. J., Major S. K., Slaga T. J. Induction of dark keratinocytes by 12-O-tetradecanoylphorbol-13-acetate and mezerein as an indicator of tumor-promoting efficiency. Carcinogenesis. 1980 May;1(5):399–406. doi: 10.1093/carcin/1.5.399. [DOI] [PubMed] [Google Scholar]
  9. Luk G. D., Goodwin G., Marton L. J., Baylin S. B. Polyamines are necessary for the survival of human small-cell lung carcinoma in culture. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2355–2358. doi: 10.1073/pnas.78.4.2355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lux G. D., Marton L. J., Baylin S. B. Ornithine decarboxylase is important in intestinal mucosal maturation and recovery from injury in rats. Science. 1980 Oct 10;210(4466):195–198. doi: 10.1126/science.6774420. [DOI] [PubMed] [Google Scholar]
  11. Marton L. J., Levin V. A., Hervatin S. J., Koch-Weser J., McCann P. P., Sjoerdsma A. Potentiation of the antitumor therapeutic effects of 1,3-bis(2-chloroethyl)-1-nitrosourea by alpha-difluoromethylornithine, an ornithine decarboxylase inhibitor. Cancer Res. 1981 Nov;41(11 Pt 1):4426–4431. [PubMed] [Google Scholar]
  12. Matsushima M., Bryan G. T. Early induction of mouse urinary bladder ornithine decarboxylase activity by rodent vesical carcinogens. Cancer Res. 1980 Jun;40(6):1897–1901. [PubMed] [Google Scholar]
  13. Mufson R. A., Fischer S. M., Verma A. K., Gleason G. L., Slaga T. J., Boutwell R. K. Effects of 12-O-tetradecanoylphorbol-13-acetate and mezerein on epidermal ornithine decarboxylase activity, isoproterenol-stimulated levels of cyclic adenosine 3':5'-monophosphate, and induction of mouse skin tumors in vivo. Cancer Res. 1979 Dec;39(12):4791–4795. [PubMed] [Google Scholar]
  14. O'Brien T. G., Simsiman R. C., Boutwell R. K. Induction of the polyamine-biosynthetic enzymes in mouse epidermis by tumor-promoting agents. Cancer Res. 1975 Jul;35(7):1662–1670. [PubMed] [Google Scholar]
  15. O'Brien T. G. The induction of ornithine decarboxylase as an early, possibly obligatory, event in mouse skin carcinogenesis. Cancer Res. 1976 Jul;36(7 Pt 2):2644–2653. [PubMed] [Google Scholar]
  16. Olson J. W., Russell D. H. Prolonged induction of hepatic ornithine decarboxylase and its relation to cyclic adenosine 3':5'-monophosphate-dependent protein kinase activation after a single administration of diethylnitrosamine. Cancer Res. 1979 Aug;39(8):3074–3079. [PubMed] [Google Scholar]
  17. Prakash N. J., Schechter P. J., Grove J., Koch-Weser J. Effect of alpha-difluoromethylornithine, an enzyme-activated irreversible inhibitor of ornithine decarboxylase, on L1210 leukemia in mice. Cancer Res. 1978 Sep;38(9):3059–3062. [PubMed] [Google Scholar]
  18. Prakash N. J., Schechter P. J., Mamont P. S., Grove J., Koch-Weser J., Sjoerdsma A. Inhibition of EMT6 tumor growth by interference with polyamine biosynthesis; effects of alpha-difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase. Life Sci. 1980 Jan 21;26(3):181–194. doi: 10.1016/0024-3205(80)90292-1. [DOI] [PubMed] [Google Scholar]
  19. Raick A. N. Cell differentiation and tumor-promoting action in skin carcinogenesis. Cancer Res. 1974 Nov;34(11):2915–2925. [PubMed] [Google Scholar]
  20. Raick A. N. Cell proliferation and promoting action in skin carcinogenesis. Cancer Res. 1974 May;34(5):920–926. [PubMed] [Google Scholar]
  21. Raick A. N. Ultrastructural, histological, and biochemical alterations produced by 12-O-tetradecanoyl-phorbol-13-acetate on mouse epidermis and their relevance to skin tumor promotion. Cancer Res. 1973 Feb;33(2):269–286. [PubMed] [Google Scholar]
  22. Seppänen P., Alhonen-Hongisto L., Jänne J. Polyamine deprivation-induced enhanced uptake of methylglyoxal bis(guanylhydrazone) by tumor cells. Biochim Biophys Acta. 1981 May 5;674(2):169–177. doi: 10.1016/0304-4165(81)90375-5. [DOI] [PubMed] [Google Scholar]
  23. Slaga T. J., Fischer S. M., Nelson K., Gleason G. L. Studies on the mechanism of skin tumor promotion: evidence for several stages in promotion. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3659–3663. doi: 10.1073/pnas.77.6.3659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Slaga T. J., Fischer S. M., Weeks C. E., Klein-Szanto A. J., Reiners J. Studies on the mechanisms involved in multistage carcinogenesis in mouse skin. J Cell Biochem. 1982;18(1):99–119. doi: 10.1002/jcb.1982.240180109. [DOI] [PubMed] [Google Scholar]
  25. Slaga T. J., Klein-Szanto A. J., Fischer S. M., Weeks C. E., Nelson K., Major S. Studies on mechanism of action of anti-tumor-promoting agents: their specificity in two-stage promotion. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2251–2254. doi: 10.1073/pnas.77.4.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Takigawa M., Inoue H., Gohda E., Asada A., Takeda Y., Mori Y. The role of putrescine in cell proliferation of the skin of mice induced by ethylphenylpropriolate. Exp Mol Pathol. 1977 Oct;27(2):183–196. doi: 10.1016/0014-4800(77)90029-6. [DOI] [PubMed] [Google Scholar]
  27. Takigawa M., Ishida H., Takano T., Suzuki F. Polyamine and differentiation: induction of ornithine decarboxylase by parathyroid hormone is a good marker of differentiated chondrocytes. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1481–1485. doi: 10.1073/pnas.77.3.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Verma A. K., Ashendel C. L., Boutwell R. K. Inhibition by prostaglandin synthesis inhibitors of the induction of epidermal ornithine decarboxylase activity, the accumulation of prostaglandins, and tumor promotion caused by 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1980 Feb;40(2):308–315. [PubMed] [Google Scholar]
  29. Verma A. K., Shapas B. G., Rice H. M., Boutwell R. K. Correlation of the inhibition by retinoids of tumor promoter-induced mouse epidermal ornithine decarboxylase activity and of skin tumor promotion. Cancer Res. 1979 Feb;39(2 Pt 1):419–425. [PubMed] [Google Scholar]
  30. Weeks C. E., Slaga T. J., Hennings H., Gleason G. L., Bracken W. M. Inhibition of phorbol ester-induced tumor promotion in mice by vitamin A analog and anti-inflammatory steroid. J Natl Cancer Inst. 1979 Aug;63(2):401–406. [PubMed] [Google Scholar]
  31. Weeks C. E., Slaga T. J. Inhibition of phorbol ester-induced polyamine accumulation in mouse epidermis by anti-inflammatory steroid. Biochem Biophys Res Commun. 1979 Dec 28;91(4):1488–1496. doi: 10.1016/0006-291x(79)91233-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES