Skip to main content
NCBI home page
Journal of Cancer Research and Clinical Oncology logoLink to Journal of Cancer Research and Clinical Oncology
. 1995 Jul;121(7):402–406. doi: 10.1007/BF01212946

Antioxidant enzymes are elevated in dimethylbenz[a]anthracene-induced neoplastic murine keratinocytes containing an activeras Ha oncogene

Kirsi Lehtola 1, Leena Laurikainen 2, Lasse Leino 1, Markku Ahotupa 3, Kari Punnonen 1,✉
PMCID: PMC12200380  PMID: 7635869

Abstract

Antioxidant enzyme activities and peroxidation potential were measured in primary mouse keratinocytes and neoplastic keratinocytes containing an activeras Ha oncogene. In neoplastic cell lines, SP-1 and 308, the activities of Cu, Zh-superoxide dismutase, catalase, and glutathione transferase were significantly elevated. The peroxidation potential was lower in cell homogenates prepared from neoplastic keratinocytes than in those prepared from normal keratinocytes. Consistently, the neoplastic 308 cell line was found to be more resistant than the normal keratinocytes to cytotoxicity induced by UV-B irradiation. The present study suggests that the enhanced antioxidant defense system protects the initiated cells from UV-B-induced oxidative stress, and that the enhanced enzymic anti-oxidant defense system is potentially a mechanism favoring the selective growth of neoplastic keratinocytes.

Key words: Antioxidants, rasHa oncogene, Keratinocyte, UV irradiation

References

  1. Beers B, Sizer W (1952) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–139 [PubMed] [Google Scholar]
  2. Cerutti PA (1995) Prooxidant states and tumor promotion. Science 227:375–381 [DOI] [PubMed] [Google Scholar]
  3. Cerutti PA (1991) Oxidant stress and carcinogenesis. Eur J Clin Invest 21:1–5 [DOI] [PubMed] [Google Scholar]
  4. Crawford DR, Amstad P, Yin Foo D, Cerutti PA (1989) Constitutive and phorbol-myristate-acetate regulated antioxidant defence of mouse epidermal JB6 cells. Mol Carcinog 2:136–143 [DOI] [PubMed] [Google Scholar]
  5. Flecha BG, Llesuy S, Boveris A (1991) Hydroperoxide-initiated chemiluminescence: an assay for oxidative stress in biopsies of heart, liver, and muscle. Free Radical Biol Med 10:93–100 [DOI] [PubMed] [Google Scholar]
  6. Glock GE, McLean P (1953) Further studies on the properties and assay of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase of rat liver. Biochem J 55:400–408 [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Habig WH, Pabst MJ, Jacoby WB (1974) Glutathione transferase. The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139 [PubMed] [Google Scholar]
  8. Halliwell B, Gutteridge JM, Cross CE (1992) Free radicals, anti-oxidants, and human disease: where are we now? J Lab Clin Med 119:598–620 [PubMed] [Google Scholar]
  9. Hartley JA, Gibson NW, Kilkenny A, Yuspa SH (1987) Mouse keratinocytes derived from initiated skin or papillomas are resistant to DNA strand breakage by benzoyl peroxide: a possible mechanism for tumor promotion mediated by benzoyl peroxide. Carcinogenesis 8:1827–1830 [DOI] [PubMed] [Google Scholar]
  10. Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH (1980) Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19:245–254 [DOI] [PubMed] [Google Scholar]
  11. Hirose K, Longo DL, Oppenheim JJ, Matsushima K (1993) Overexpression of mitochondrial manganese superoxide dismutase promotes the survival of tumor cells exposed to interleukin-1, tumor necrosis factor, selected anticancer drugs, and ionizing radiation. FASEB J 7:361–368 [DOI] [PubMed] [Google Scholar]
  12. Kobayashi T, Matsumoto M, Iizuka H, Suzuki K, Taniguchi N (1991) Superoxide dismutase in psoriasis, squamous cell carcinoma and basal cell epithelioma: an immunohistochemical study. Br J Dermatol 124:555–559 [DOI] [PubMed] [Google Scholar]
  13. Laihia JK, Jansen CT, Ahotupa M (1993) Lucigenin and linoleate enhanced chemiluminescent assay for superoxide dismutase activity. Free Radical Biol Med 14:457–461 [DOI] [PubMed] [Google Scholar]
  14. Layne EK (1957) Spectrophotometric and turbidimetric methods for measuring protein. Methods Enzymol 3:447–454 [Google Scholar]
  15. Marklund S, Westman N, Lundgren E, Roos G (1982) Copper-and zinc-containing superoxide dismutase, manganese-containing superoxide dismutase, catalase, and glutathione peroxidase in normal and neoplastic human cell lines and normal human tissues. Cancer Res 42:1955–1961 [PubMed] [Google Scholar]
  16. Nishi J, Ogura R, Sugiyama M, Hidaka T, Kohno M (1991) involvement of active oxygen in lipid peroxide radical reaction of epidermal homogenate following ultraviolet light exposure. J Invest Dermatol 97:115–119 [DOI] [PubMed] [Google Scholar]
  17. Paglia D, Valentine W (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70:158–169 [PubMed] [Google Scholar]
  18. Punnonen K, Yuspa SH (1992) Ultraviolet light irradiation increases cellular diacylglycerol and induces translocation of diacylglycerol kinase in murine keratinocytes. J Invest Dermatol 99:221–226 [DOI] [PubMed] [Google Scholar]
  19. Punnonen K, Puustinen T, Jansen CT (1987) The effect of UVB irradiation on distribution and release of dihomo-γ-linolenic acid, arachidonic acid, and eicosapentaenoic acid in human keratinocytes in culture. J Invest Dermatol 88:611–614 [DOI] [PubMed] [Google Scholar]
  20. Punnonen K, Autio P, Kiistala U, Ahotupa M (1991a) In-vivo effects of solar-simulated ultraviolet irradiation on antioxidant enzymes and lipid peroxidation in human epidermis. Br J Dermatol 125:18–20 [DOI] [PubMed] [Google Scholar]
  21. Punnonen K, Jansen CT, Puntala A, Ahotupa M (1991b) Effects of in vitro UVA irradiation and PUVA treatment on membrane fatty acids and activities of antioxidant enzymes in human-keratinocytes. J Invest Dermatol 96:255–259 [DOI] [PubMed] [Google Scholar]
  22. Punnonen K, Puntala A, Ahotupa M (1991c) Effects of ultraviolet A and B irradiation on lipid peroxidation and activity of the antioxidant enzymes in keratinocytes in culture. Photodermatol Photoimmunol Photomed 8:3–6 [PubMed] [Google Scholar]
  23. Punnonen K, Puntala A, Jansen CT, Ahotupa M (1991d) UVB irradiation induces lipid peroxidation and reduces antioxidant enzyme activities in human keratinocytes in vitro. Acta Derm Venereol (Stockh) 71:239–273 [PubMed] [Google Scholar]
  24. Punnonen R, Kudo R, Punnonen K, Hietanen E, Kuoppala T, Kainulainen H, Sato K, Ahotupa M (1993) Activities of anti-oxidant enzymes and lipid peroxidation in endometrial cancer. Eur J Cancer 29A: 266–269 [DOI] [PubMed] [Google Scholar]
  25. Shindo Y, Witt E, Packer L (1993) Antioxidant defence mechanisms in murine epidermis and dermis and their responses to ultraviolet light. J Invest Dermatol 100:260–265 [DOI] [PubMed] [Google Scholar]
  26. Sies H (1985) Oxidative stress: Introductory remarks. In: Sies H (ed) Oxidative stress. Academic Press, New York, pp 1–8 [Google Scholar]
  27. Strickland JE, Greenhalgh DA, Koceva-Chyla A, Hennings H, Restrepo C, Balaschak, Yuspa SH (1988) Development of murine epidermal cell lines which contain an activated rasHa oncogene and form papillomas in skin grafts on athymic nude mouse hosts. Cancer Res 48:165–169 [PubMed] [Google Scholar]
  28. Sun Y (1990) Free radicals, antioxidant enzymes, and carcinogenesis. Free Radical Biol Med 8:583–599 [DOI] [PubMed] [Google Scholar]
  29. Sun Y, Oberley LW, Oberley TD, Elwell JH, Sierra-Rivera E (1993) Lowered antioxidant enzymes in spontaneously transformed embryonic mouse liver cells in culture. Carcinogenesis 14:1457–1463 [DOI] [PubMed] [Google Scholar]
  30. Yuspa SH, Kilkenny A, Cheng C, Roop D, Hennings H, Kruszewski F, Lee E, Strickland J, Greenhalgh DA (1991) Alterations in epidermal biochemistry as a consequence of stage-specific genetic changes in skin carcinogenesis. Environ Health Perspect 93:3–10 [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yuspa SH, Punnonen K, Lee E, Hennings H, Strickland JE, Cheng C, Glick A, Dlugosz A (1992) Alterations in the biology and biochemistry of keratinocytes induced by theras oncogene. Prog Clin Biol Res 376:103–115 [PubMed] [Google Scholar]

Articles from Journal of Cancer Research and Clinical Oncology are provided here courtesy of Springer

RESOURCES