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. 1991 Feb 1;88(3):946–950. doi: 10.1073/pnas.88.3.946

Free radical-derived quinone methide mediates skin tumor promotion by butylated hydroxytoluene hydroperoxide: expanded role for electrophiles in multistage carcinogenesis.

K Z Guyton 1, P Bhan 1, P Kuppusamy 1, J L Zweier 1, M A Trush 1, T W Kensler 1
PMCID: PMC50931  PMID: 1846971

Abstract

Free radical derivatives of peroxides, hydroperoxides, and anthrones are thought to mediate tumor promotion by these compounds. Further, the promoting activity of phorbol esters is attributed, in part, to their ability to stimulate the cellular generation of oxygen radicals. A hydroperoxide metabolite of butylated hydroxytoluene, 2,6-di-tert-butyl-4-hydroperoxyl-4-methyl-2,5-cyclohexadienone (BHTOOH), has previously been shown to be a tumor promoter in mouse skin. BHTOOH is extensively metabolized by murine keratinocytes to several radical species. The primary radical generated from BHTOOH is a phenoxyl radical that can disproportionate to form butylated hydroxytoluene quinone methide, a reactive electrophile. Since electrophilic species have not been previously postulated to mediate tumor promotion, the present study was undertaken to examine the role of this electrophile in the promoting activity of BHTOOH. The biological activities of two chemical analogs of BHTOOH, 4-trideuteromethyl-BHTOOH and 4-tert-butyl-BHTOOH, were compared with that of the parent compound. 4-Trideuteromethyl-BHTOOH and 4-tert-butyl-BHTOOH have a reduced ability or inability, respectively, to form a quinone methide; however, like the parent compound, they both generate a phenoxyl radical when incubated with keratinocyte cytosol. The potency of BHTOOH, 4-trideuteromethyl-BHTOOH, and 4-tert-butyl-BHTOOH as inducers of ornithine decarboxylase, a marker of tumor promotion, was commensurate with their capacity for generating butylated hydroxytoluene quinone methide. These initial results were confirmed in a two-stage tumor promotion protocol in female SENCAR mice. Together, these data indicate that a quinone methide is mediating tumor promotion by BHTOOH, providing direct evidence that an electrophilic intermediate can elicit this stage of carcinogenesis.

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Selected References

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  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Cerutti P. A. Prooxidant states and tumor promotion. Science. 1985 Jan 25;227(4685):375–381. doi: 10.1126/science.2981433. [DOI] [PubMed] [Google Scholar]
  3. Cohen S. M., Ellwein L. B. Cell proliferation in carcinogenesis. Science. 1990 Aug 31;249(4972):1007–1011. doi: 10.1126/science.2204108. [DOI] [PubMed] [Google Scholar]
  4. DiGiovanni J., Kruszewski F. H., Coombs M. M., Bhatt T. S., Pezeshk A. Structure-activity relationships for epidermal ornithine decarboxylase induction and skin tumor promotion by anthrones. Carcinogenesis. 1988 Aug;9(8):1437–1443. doi: 10.1093/carcin/9.8.1437. [DOI] [PubMed] [Google Scholar]
  5. Farber E. The multistep nature of cancer development. Cancer Res. 1984 Oct;44(10):4217–4223. [PubMed] [Google Scholar]
  6. Inai K., Kobuke T., Nambu S., Takemoto T., Kou E., Nishina H., Fujihara M., Yonehara S., Suehiro S., Tsuya T. Hepatocellular tumorigenicity of butylated hydroxytoluene administered orally to B6C3F1 mice. Jpn J Cancer Res. 1988 Jan;79(1):49–58. doi: 10.1111/j.1349-7006.1988.tb00010.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ito N., Fukushima S., Tsuda H. Carcinogenicity and modification of the carcinogenic response by BHA, BHT, and other antioxidants. Crit Rev Toxicol. 1985;15(2):109–150. doi: 10.3109/10408448509029322. [DOI] [PubMed] [Google Scholar]
  8. Kahl R. Synthetic antioxidants: biochemical actions and interference with radiation, toxic compounds, chemical mutagens and chemical carcinogens. Toxicology. 1984 Dec;33(3-4):185–228. doi: 10.1016/0300-483x(84)90038-6. [DOI] [PubMed] [Google Scholar]
  9. Kalyanaraman B., Mottley C., Mason R. P. A direct electron spin resonance and spin-trapping investigation of peroxyl free radical formation by hematin/hydroperoxide systems. J Biol Chem. 1983 Mar 25;258(6):3855–3858. [PubMed] [Google Scholar]
  10. Kozumbo W. J., Seed J. L., Kensler T. W. Inhibition by 2(3)-tert-butyl-4-hydroxyanisole and other antioxidants of epidermal ornithine decarboxylase activity induced by 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1983 Jun;43(6):2555–2559. [PubMed] [Google Scholar]
  11. Malkinson A. M., Beer D. S. Pharmacologic and genetic studies on the modulatory effects of butylated hydroxytoluene on mouse lung adenoma formation. J Natl Cancer Inst. 1984 Oct;73(4):925–933. [PubMed] [Google Scholar]
  12. Malkinson A. M., Thaete L. G., Blumenthal E. J., Thompson J. A. Evidence for a role of tert-butyl hydroxylation in the induction of pneumotoxicity in mice by butylated hydroxytoluene. Toxicol Appl Pharmacol. 1989 Nov;101(2):196–204. doi: 10.1016/0041-008x(89)90269-x. [DOI] [PubMed] [Google Scholar]
  13. Masuda Y., Nakayama N. Prevention of butylated hydroxytoluene-induced lung damage by diethyldithiocarbamate and carbon disulfide in mice. Toxicol Appl Pharmacol. 1984 Aug;75(1):81–90. doi: 10.1016/0041-008x(84)90078-4. [DOI] [PubMed] [Google Scholar]
  14. Miller J. A. Carcinogenesis by chemicals: an overview--G. H. A. Clowes memorial lecture. Cancer Res. 1970 Mar;30(3):559–576. [PubMed] [Google Scholar]
  15. Mizutani T., Nomura H., Nakanishi K., Fujita S. Hepatotoxicity of butylated hydroxytoluene and its analogs in mice depleted of hepatic glutathione. Toxicol Appl Pharmacol. 1987 Jan;87(1):166–176. doi: 10.1016/0041-008x(87)90094-9. [DOI] [PubMed] [Google Scholar]
  16. Mizutani T., Nomura H., Yamamoto K., Tajima K. Modification of butylated hydroxytoluene-induced pulmonary toxicity in mice by diethyl maleate, buthionine sulfoximine, and cysteine. Toxicol Lett. 1984 Dec;23(3):327–331. doi: 10.1016/0378-4274(84)90029-8. [DOI] [PubMed] [Google Scholar]
  17. Mizutani T., Yamamoto K., Tajima K. Isotope effects on the metabolism and pulmonary toxicity of butylated hydroxytoluene in mice by deuteration of the 4-methyl group. Toxicol Appl Pharmacol. 1983 Jun 30;69(2):283–290. doi: 10.1016/0041-008x(83)90310-1. [DOI] [PubMed] [Google Scholar]
  18. Mizutani T., Yamamoto K., Tajima K. Isotope effects on the metabolism and pulmonary toxicity of butylated hydroxytoluene in mice by deuteration of the 4-methyl group. Toxicol Appl Pharmacol. 1983 Jun 30;69(2):283–290. doi: 10.1016/0041-008x(83)90310-1. [DOI] [PubMed] [Google Scholar]
  19. O'Brien T. G., Simsiman R. C., Boutwell R. K. Induction of the polyamine-biosynthetic enzymes in mouse epidermis and their specificity for tumor promotion. Cancer Res. 1975 Sep;35(9):2426–2433. [PubMed] [Google Scholar]
  20. Olsen P., Meyer O., Bille N., Würtzen G. Carcinogenicity study on butylated hydroxytoluene (BHT) in Wistar rats exposed in utero. Food Chem Toxicol. 1986 Jan;24(1):1–12. doi: 10.1016/0278-6915(86)90256-5. [DOI] [PubMed] [Google Scholar]
  21. Taffe B. G., Kensler T. W. Tumor promotion by a hydroperoxide metabolite of butylated hydroxytoluene, 2,6-di-tert-butyl-4-hydroperoxy-4-methyl-2,5-cyclohexadienone, in mouse skin. Res Commun Chem Pathol Pharmacol. 1988 Sep;61(3):291–303. [PubMed] [Google Scholar]
  22. Taffe B. G., Takahashi N., Kensler T. W., Mason R. P. Generation of free radicals from organic hydroperoxide tumor promoters in isolated mouse keratinocytes. Formation of alkyl and alkoxyl radicals from tert-butyl hydroperoxide and cumene hydroperoxide. J Biol Chem. 1987 Sep 5;262(25):12143–12149. [PubMed] [Google Scholar]
  23. Taffe B. G., Zweier J. L., Pannell L. K., Kensler T. W. Generation of reactive intermediates from the tumor promoter butylated hydroxytoluene hydroperoxide in isolated murine keratinocytes or by hematin. Carcinogenesis. 1989 Jul;10(7):1261–1268. doi: 10.1093/carcin/10.7.1261. [DOI] [PubMed] [Google Scholar]
  24. Thompson J. A., Schullek K. M., Fernandez C. A., Malkinson A. M. A metabolite of butylated hydroxytoluene with potent tumor-promoting activity in mouse lung. Carcinogenesis. 1989 Apr;10(4):773–775. doi: 10.1093/carcin/10.4.773. [DOI] [PubMed] [Google Scholar]
  25. Valoti M., Sipe H. J., Jr, Sgaragli G., Mason R. P. Free radical intermediates during peroxidase oxidation of 2-t-butyl-4-methoxyphenol, 2,6-di-t-butyl-4-methylphenol, and related phenol compounds. Arch Biochem Biophys. 1989 Mar;269(2):423–432. doi: 10.1016/0003-9861(89)90126-4. [DOI] [PubMed] [Google Scholar]
  26. Wand M. D., Thompson J. A. Cytochrome P-450-catalyzed rearrangement of a peroxyquinol derived from butylated hydroxytoluene. Involvement of radical and cationic intermediates. J Biol Chem. 1986 Oct 25;261(30):14049–14056. [PubMed] [Google Scholar]
  27. Witschi H., Malkinson A. M., Thompson J. A. Metabolism and pulmonary toxicity of butylated hydroxytoluene (BHT). Pharmacol Ther. 1989;42(1):89–113. doi: 10.1016/0163-7258(89)90023-5. [DOI] [PubMed] [Google Scholar]
  28. Yamamoto K., Tajima K., Mizutani T. The acute toxicity of butylated hydroxytoluene and its metabolites in mice. Toxicol Lett. 1980 Aug;6(3):173–175. doi: 10.1016/0378-4274(80)90187-3. [DOI] [PubMed] [Google Scholar]
  29. Yumibe N. P., Thompson J. A. Fate of free radicals generated during one-electron reductions of 4-alkyl-1,4-peroxyquinols by cytochrome P-450. Chem Res Toxicol. 1988 Nov-Dec;1(6):385–390. doi: 10.1021/tx00006a010. [DOI] [PubMed] [Google Scholar]

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