Abstract
Although many studies have been reported on the repair of ultraviolet light (UV)-induced cyclobutane-type pyrimidine dimers (CPDs) in DNA, the effects of aging on the removal of UV-induced CPDs from the human skin epidermis in vivo remains uncertain. Therefore, we employed immunoblotting and immunohistochemical methods using monoclonal antibodies (TDM-2) to CPDs to study age-related differences in the time required for the in vivo removal of UVB-induced CPDs. The flexure surfaces of the upper arms of five young men were exposed to UVB light at a fluence of 35 and 700 mJ/cm2, and four older men were also irradiated with the same doses of UVB mentioned above. Each area of skin was biopsied before and immediately after irradiation, and at 4, 24 h, 2 and 4 days after irradiation in the younger group; and before and immediately after irradiation, and at 24 h, 4, 7, and 14 days after irradiation in the older group. A total of 108 DNA samples were taken from the epidermis of 108 biopsied specimens. These samples were immunoblotted using TDM-2 and the intensities of the immunoprecipitates were measured by photodensitometer. Our results show that the CPDs had been removed from the epidermis at 4 days after irradiation at either dose in the younger group, and between 7–14 days after irradiation in the aged group. The results of our immunohistochemical studies were consistent with those of our immunoblotting studies, and indicated that basal cells repair CPDs more quickly than prickle cells in the epidermis except the amounts at 24 h after UVB irradiation, and that the CPDs were removed by epidermal turnover after the nucleotide excision repair (NER). Our results showed age-associated decline in the NER in vivo, indicating high risk of UV-associated skin cancer.
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References
Alcalay J, Freeman SE, Goldberg LH, Wolf JE (1990) Excision repair of pyrimidine dimers induced by simulated solar radiation in the skin of patients with basal cell carcinoma. J Invest Dermatol 95:506–509
Baden HP, Pearlman C (1964) The effects of ultraviolet light on protein and nucleic acid synthesis in the epidermis. J Invest Dermatol 48:71–75
Balmain A (1985) Transforming ras oncogenes and multistage carcinogenesis. Br J Cancer 51:1–7
Beukers R, Berend SW (1961) The effects of UV-irradiation on nucleic acid and their components. Biochem Biophys Acta 49:181–189
Blin N, Stafford DW (1976) A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res 3:2303
Boer J, Hoeijmakers HJ (2000) Nucleotide excision repair and human syndromes. Carcinogenesis 21:453–460
Boyd AS, Stasko T, Cameron GS, Russell M, King LE (2001) Histologic features of actinic keratosis in solid organ transplant recipients and healthy controls. J Am Acad Dermatol 45:217–21
Christiansen M, Stevnsner T, Bohr VA, Clark BFC, Rattan SIS (2000) Gene-specific DNA repair of pyrimidine dimers does not decline during cellular aging in vitro. Exp Cell Res 256:308–314
Cleaver JE (1968) Defective repair of DNA in xeroderma pigmentosum. Nature 218:652–656
D’Ambrosio SM, Slazinski L, Whetstone JW, Lowney E (1981) Excision repair of UV-induced pyrimidine dimers in human skin in vivo. J Invest Dermatol 77:311–313
Eggset G, Volden G, Krokan H (1983) UV-induced DNA damage and its repair in human skin in vivo studied by sensitive immunohistochemical method. Carcinogenesis 4:745–750
Epstein JH (1970) Effects of ultraviolet radiation on the mitotic cycle and DNA, RNA and protein synthesis in mammalian epidermis in vivo. Photochem Photobiol 12:57–65
Epstein WL, Fukuyama K, Epstein JH (1971) Ultraviolet light, DNA repair and skin carcinogenesis in man. Fed Proc 30:1766–1771
FranklinWA, HaseltineWA (1984) Removal of UV-light-induced pyrimidine-pyrimidon (6–4) products for Escherichia coli DNA requires the uvrA, uvrB and uvrC gene products. Proc Natl Acad Sci USA 81:3821–3824
Freeman SE (1988) Variation in excision repair of UVB-induced pyrimidine dimers in DNA of human skin in situ. Invest Dermatol 90:814–817
Goukassian D, Gad F, Yaar M, Eller MS, Nehal US, Gilchrest BA (2000) Mechanisms and implications of the age-associated decrease in DNA repair capacity. FASEB J 14:1325–1334
Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ (1993) The p21 Cdk-interacting protein Cipl is a potent inhibitor of G1 cyclin-dependent kinase. Cell 75:805–816
Hori M, Udono UM, Yoshida H, Urata Y, Koike K, Ihara M (1992) Immunochemical detection of unrepaired cyclobutane-type pyrimidine dimers of DNAs extracted from human skin tumours. Arch Dermatol Res 284:283–289
Lambert B, Ringbord U, Swanbeck G (1978) Ultraviolet-induced DNA repair synthesis in lymphocytes from patients with actinic keratosis. J Invest Dermatol 67:594–598
Lane DP (1992) p53, guardian of the genome. Nature 358:15–16
Lond M, Parada LF, Weinberg RA (1983) Cellular oncogene and multistep carcinogenesis. Science 222:771–778
Matunaga T, Mori T, Nikaido O (1990) Base sequence specificity of monoclonal antibody to (6–4) photoproducts. Mutat Res 235:187–194
McDonell M, Simon MN, Studier FW (1977) Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis of neutral and alkaline gels. J Mol Biol 110:119–143
Miller JH (1985) Mutagenic specificity of ultraviolet-light. J Mol Biol 182:45–68
Mizuno T, Matunaga N, Ihara M, Nikaido O (1991) Establishment of monoclonal antibody recognizing cyclobutane-type thymine dimers in DNA: comparative study with 64M-1 antibody specific for (6–4) photoproducts. Mutat Res 254:175–184
Mori T, Matunaga T, Hirose T, Nikaido O (1988) Establishment of a monoclonal antibody recognizing ultraviolet light-induced (6–4) photoproducts. Mutat Res 194:263–270
Mori T, Nakane M, Hattori T, Matunaga T, Ihara M, Nikaido O (1991) Simultaneous establishment of monoclonal antibodies specific for either cyclobutane pyrimidine dimer or (6–4) photoproduct from the same mouse immunized with ultraviolet-irradiated DNA. Photochem Photobiol 54:225–232
Mori M, Kobayashi H, Sugiyama C, Katsumura Y, Furihata C (2000) Effect of aging on unscheduled DNA synthesis induction by UV-B irradiation in hairless mouse epidermis. J Toxicol Sci 25:181–188
Moriwaki S, Ray S, Tarone RE, Kraemer KH, Grossman L (1996) The effect of donor age on the processing of UV-damaged DNA by cultured human cells: reduced DNA repair capacity and increased DNA mutability. Mutat Res 364:117–123
Patric MH (1970) Near-UV photolysis of pyrimidine heteroadducts in E. coli DNA. Photochem Photobiol 447–485
Patric MH (1977) Studies thymine-derived photoproducts in DNA I. Formation and biological role of pyrimidine adducts in DNA. Photochem Photobiol 25:357–372
Price ML, Tidman MJ, Fagg NLK, PalmerTJ, MacDonald DM (1988) Distinctive epidermal atypia in immunosuppression-associated cutaneous malignancy. Histopathology 13:89–94
Protic-Sabljic M, Tuteja N, Munsen P, Hauser J, Kraemer KH, Dixon K (1986) UV-induced cyclobutane pyrimidine dimers are mutagenic in mammalian cells. Mol Cell Biol 6:3349–3356
Qin X, Zhang S, Nakatsuru Y, Oda H, Yamazaki Y, Suzuki T, Nikaido O, Ishikawa T (1994) Detection of active UV-photoproduct repair in monkey skin in vivo by quantitative immunohistochemistry. Cancer Lett 83:291–298
Reusch MK, Meager K, Leadon SA, Hanawalt PC (1988) Comparative remove 1 of pyrimidine dimers from human epidermal karatinocyte in vivo and vitro. J Invest Dermatol 91:349–352
Rosa LK, Vander Wulp IM, MacFarlane SJ, Lohman PHM, Baan RA (1988) Detection of cyclobutane thymine dimers in DNA of human cells with monoclonal antibodies raised against a thymine dimer containing tetranucleotide. Photochem Photobiol 48:627–633
Sancar GB, Siede W, van Zeeland AA (1996) Repair and processing of DNA damage: a summary of recent progress. Mutat Res 362:127–146
Sbano E, Andreassi M, Valentino A, Baiocchi R (1978) DNA-repair after UV-irradiation in skin fibroblasts from patients with actinic keratosis. Arch Dermatol Res 262:55–61
Setlow RB (1966) Cyclobutane-type pyrimidine dimers in polynucleotides. Science 153:379–380
Strickland PT, Boyle JM (1981) Characterization of two monoclonal antibodies specific for dimerized and nondimerized adjacent thymine in single DNA. Photochem Photobiol 34:595–601
Sutherland BM, Harber LC, Kochevar IE (1980) Pyrimidine dimer formation and repair in human skin. Cancer Res 40:3181–3185
Suzuki F, Han A, Lankas GR, Utsumi H, Elkind MM (1981) Spectral dependencies of killing, mutation and transformation in mammalian cells and their relevance to hazards caused by solar ultraviolet radiation. Cancer Res 41:4916–4924
Udono UM, Hori M, Hirayama M, Yoshida H, Suematsu T, Nikaido O (1993) Visualization of ultraviolet light-induced thymine dimers in DNA by immunoelectron microscopy. Photochem Photobiol 57:752–754
Varghese AJ, Wand SY (1967) Cis-syn thymine homodimer from ultraviolet irradiated calf thymus DNA. Nature 213:909–910
Varghese AJ, Wand SY (1968) Thymine–thymine adduct as a photoproduct of thymine. Science 160:186–187
Wani AA, Ambrosio SMD, Alvi NK (1987) Quantitation of pyrimidine dimers by immunoslot blot following subleathal UV-irradiation of human cells. Photochem Photobiol 46:477–482
Wani AA, Gibson D’Ambrosio RE, D’Ambrosio SM (1984) Antibodies to UV-irradiated DNA: the monitoring of DNA danage by ELISA and indirect immunofluorescence. Photochem Photobiol 40:465–471
Wei Q, Matanoski GM, Farmer ER, Hedayati MA, Grossman L (1995) DNA repair capacity for ultraviolet light-induced damage is reduced in peripheral lymphocytes from patients with basal cell carcinoma. J Invest Dermatol 104:933–936
Wei Q (1998) Effect of aging on DNA repair and skin carcinogenesis: a minireview of population-based studies. J Invest Dermatol Symp Proc 3:19–22
Wood RD, Skopek TR, Huchinson F (1986) Changes in DNA base sequence by targeted mutagenesis of lambda phage by ultraviolet light. J Mol Biol 173:273–291
Wood RD (1996) DNA repair in eukaryotes. Annu Rev Biochem 65:135–167
Xu G, Snellman E, Bykov VJ, Jansen CT, Hemminki K (2000) Effect of age on the formation and repair of UV photoproducts in human skin in situ. Mutat Res 459:195–202
Yarosh D (2004) DNA repair, immunosuppression, and skin cancer. Cutis. 74 (Suppl 5):10–13
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Yamada, M., Udono, M.U., Hori, M. et al. Aged human skin removes UVB-induced pyrimidine dimers from the epidermis more slowly than younger adult skin in vivo. Arch Dermatol Res 297, 294–302 (2006). https://doi.org/10.1007/s00403-005-0618-0
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DOI: https://doi.org/10.1007/s00403-005-0618-0