Abstract
Mechanisms of UVA-mutagenesis remain a matter of debate. Earlier described higher rates of mutation formation per pyrimidine dimer with UVA than with UVB and other evidence suggested that a non-pyrimidine dimer-type of DNA damage contributes more to UVA- than to UVB-mutagenesis. However, more recently published data on the spectra of UVA-induced mutations in primary human skin cells and in mice suggest that pyrimidine dimers are the most common type of DNA damage-inducing mutations not only with UVB, but also with UVA. As this rebuts a prominent role of non-dimer type of DNA damage in UVA-mutagenesis, we hypothesized that the higher mutation rate at UVA-induced pyrimidine dimers, as compared to UVB-induced ones, is caused by differences in the way UVA- and UVB-exposed cells process DNA damage. Therefore, we here compared cell cycle regulation, DNA repair, and apoptosis in primary human fibroblasts following UVB- and UVA-irradiation, using the same physiologic and roughly equimutagenic doses (100–300 J m−2 UVB, 100–300 kJ m−2 UVA) we have used previously for mutagenesis experiments with the same type of cells. ELISAs for the detection of pyrimidine dimers confirmed that much fewer dimers were formed with these doses of UVA, as compared to UVB. We found that cell cycle arrests (intra-S, G1/S, G2/M), mediated at least in part by activation of p53 and p95, are much more prominent and long-lasting with UVB than with UVA. In contrast, no prominent differences were found between UVA and UVB for other anti-mutagenic cellular responses (DNA repair, apoptosis). Our data suggest that less effective anti-mutagenic cellular responses, in particular different and shorter-lived cell cycle arrests, render pyrimidine dimers induced by UVA more mutagenic than pyrimidine dimers induced by UVB.
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E. El Ghissassi, R. Baan, K. Straif, Y. Grosse, B. Secretan, V. Bouvard, L. Benbrahim-Talla, N. Guha, C. Freeman, L. Galichet, V. Cogliano, and on behalf of the WHO International Agency for Research on Cancer Monograph Working Group, A review of human carcinogens - part D: radiation, Lancet Oncol., 2009, 10, 751–752.
J. Cadet, E. Sage, T. Douki, Ultraviolet radiation-mediated damage to cellular DNA, Mutat. Res., Fundam. Mol. Mech. Mutagen., 2005, 571, 3–17.
C. Kielbassa, L. Roza, B. Epe, Wavelength dependence of oxidative DNA damage induced by UV and visible light, Carcinogenesis, 1997, 18, 811–816.
S. E. Freeman, R. W. Gange, J. C. Sutherland, E. A. Matzinger, B. M. Sutherland, Production of pyrimidine dimers in DNA of human skin exposed in situ to UVA radiation, J. Invest. Dermatol., 1987, 88, 430–433.
T. Matsunaga, K. Hieda, O. Nikaido, Wavelength dependent formation of thymine dimers and (6–4) photoproducts in DNA by monochromatic ultraviolet light ranging from 150 to 365 nm, Photochem. Photobiol., 1991, 54, 403–410.
R. D. Ley, A. Fourtanier, UVA1-induced edema and pyrimidine dimers in murine skin, Photochem. Photobiol., 2000, 72, 485–487.
A. R. Young, C. S. Potten, O. Nikaido, P. G. Parsons, J. Boenders, J. M. Ramsden, C. A. Chadwick, Human melanocytes and keratinocytes exposed to UVB or UVA in vivo show comparable levels of thymine dimers, J. Invest. Dermatol., 1998, 111, 936–940.
Z. Kuluncsics, D. Perdiz, E. Brulay, B. Muel, E. Sage, Wavelengths dependence of ultraviolet-induced DNA damage distribution: involvement of direct or indirect mechanisms and possible artefacts, J. Photochem. Photobiol., B, 1999, 49, 71–80.
T. Douki, D. Perdiz, P. Grof, Z. Kuluncsics, E. Moustacchi, J. Cadet, E. Sage, Oxidation of guanine in cellular DNA by solar UV radiation: biological role, Photochem. Photobiol., 1999, 70, 184–190.
S. Courdavault, C. Baudouin, M. Charveron, A. Favier, J. Cadet, T. Douki, Larger yield of cyclobutane dimers than 8-oxo-7,8-dihydroguanine in the DNA of UVA-irradiated human skin cells, Mutat. Res., 2004, 556, 135–142.
T. M. Rünger, U. P. Kappes, Mechanisms of mutation formation with long-wave ultraviolet light (UVA), Photodermatol., Photoimmunol. Photomed., 2008, 24, 2–10.
U. P. Kappes, D. Luo, M. Potter, K. Schulmeister, T. M. Rünger, Short- and long-wave ultraviolet light (UVB and UVA) induce similar mutations in human skin cells, J. Invest. Dermatol., 2006, 126, 667–675.
C. Robert, H. Mueller, A. Benoit, L. Dubertret, A. Sarasin, A. Stary, Cell survival and shuttle vector mutagenesis induced by ultraviolet A and ultraviolet B radiation in a human cell line, J. Invest. Dermatol., 1996, 106, 721–728.
H. Ikehata, K. Kawai, J. Komura, K. Sakatsume, L. Wang, M. Imai, S. Higashi, O. Nikaido, K. Yamamoto, K. Hieda, M. Watanabe, H. Kasai, T. Ono, UVA1 genotoxicity is meditated not by oxidative damage but by cyclobutane pyrimidine dimers in normal mouse skin, J. Invest. Dermatol., 2008, 128, 2289–2296.
I. C. Enninga, R. T. L. Groenendijk, A. R. Filon, A. A. van Zeeland, J. W. I. M. Simons, The wavelength dependence of U.V.-induced pyrimidine dimer formation, cell killing and mutation induction in human diploid skin fibroblasts, Carcinogenesis, 1986, 7, 1829–1836.
F. R. de Gruijl, H. J. Sterenborg, P. D. Forbes, R. E. Davies, C. Cole, G. Kelfkens, W. H. van, H. Slaper, J. C. van der Leun, F. R. de Gruijl, H. J. Sterenborg, P. D. Forbes, R. E. Davies, C. Cole, G. Kelfkens, H. van Weelden, H. Slaper, J. C. van der Leun, Wavelength dependence of skin cancer induction by ultraviolet irradiation of albino hairless mice, Cancer Res., 1993, 53, 53–60.
F. R. de Gruijl, Photocarcinogenesis: UVA vs. UVB, Methods Enzymol., 2000, 319, 359–366.
P. C. Hanawalt, D. J. Crowley, J. M. Ford, A. K. Ganesan, D. R. Lloyd, T. Nouspikel, C. A. Smith, G. Spivak, S. Tornaletti, Regulation of nucleotide excision repair in bacteria and mammalian cells, Cold Spring Harbor Symp. Quant. Biol., 2000, 65, 183–191.
S. Bhana, D. R. Lloyd, The role of p53 in DNA damage-mediated cytotoxicity overrides its ability to regulate nucleotide excision repair in human fibroblasts, Mutagenesis, 2008, 23, 43–50.
B. M. Stanulis-Praeger, B. A. Gilchrest, Effect of donor age and prior sun exposure on growth inhibition of cultured human dermal fibroblasts by all trans-retinoic acid, J. Cell. Physiol., 1989, 139, 116–124.
K. Werninghaus, R. M. Handjani, B. A. Gilchrest, Protective effect of alpha-tocopherol in carrier liposomes on ultraviolet-mediated human epidermal cell damage in vitro, Photodermatol., Photoimmunol. Photomed., 1991, 8, 236–242.
A. Stary, A. Sarasin, Ultraviolet A- and singlet oxygen-induced mutation spectra,, Methods Enzymol., 2000, 319, 153–165.
R. M. Sayre, C. Cole, W. Billhimer, J. Stanfield, R. D. Ley, Spectral comparison of solar simulators and sunlight, Photodermatol., Photoimmunol. Photomed., 1990, 7, 159–165.
K. M. Thoms, C. Kuschal, E. Oetjen, T. Mori, N. Kobayashi, P. Laspe, L. Boeckmann, M. P. Schon, S. Emmert, Cyclosporin A, but not everolimus, inhibits DNA repair mediated by calcineurin: implications for tumorigenesis under immunosuppression, Exp. Dermatol., 2011, 20, 232–236.
T. Mori, M. Nakane, T. Hattori, T. Matsunaga, M. Ihara, O. Nikaido, T. Mori, M. Nakane, T. Hattori, T. Matsunaga, M. Ihara, O. Nikaido, 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., 1991, 54, 225–232.
A. de Laat, M. van Tilburg, J. C. von der Leun, W. A. van Vloten, F. R. de Gruijl, Cell cycle kinetics following UVA irradiation in comparison to UVB and UVC irradiation, Photochem. Photobiol., 1996, 63, 492–497.
R. M. Snapka, Bromodeoxyuridine photodamage in studies of UVA damage and the cell cycle, DNA Repair, 2009, 8, 3.
T. M. Rünger, B. Epe, K. Möller, Repair of ultraviolet B and singlet oxygen-induced DNA damage in xeroderma pigmentosum cells, J. Invest. Dermatol., 1995, 104, 68–73.
S. Emmert, H. Slor, D. B. Busch, S. Batko, R. B. Albert, D. Coleman, S. G. Khan, B. Abu-Libdeh, J. J. DiGiovanna, B. B. Cunningham, M. M. Lee, J. Crollick, H. Inui, T. Ueda, M. Hedayati, L. Grossman, T. Shahlavi, J. E. Cleaver, K. H. Kraemer, Relationship of neurologic degeneration to genotype in three xeroderma pigmentosum group G patients, J. Invest. Dermatol., 2002, 118, 972–982.
T. M. Rünger, B. Epe, K. Möller, Processing of directly and indirectly ultraviolet-induced DNA damage in human cells, Recent Results Cancer Res., 1995, 139, 31–42.
S. Emmert, N. Kobayashi, S. G. Khan, K. H. Kraemer, The xeroderma pigmentosum group C gene leads to selective repair of cyclobutane pyrimidine dimers rather than 6-4 photoproducts, Proc. Natl. Acad. Sci. U. S. A., 2000, 97, 2151–2156.
J. Dunn, M. Potter, A. Rees, T. M. Rünger, Activation of the Fanconi anemia/BRCA pathway and recombination repair in the cellular response to solar UV, Cancer Res., 2006, 66, 11140–11147.
J. M. Ford, Regulation of DNA damage recognition and nucleotide excision repair: another role for p53, Mutat. Res., Fundam. Mol. Mech. Mutagen., 2005, 577, 195–202.
S. Adimoolam, J. M. Ford, p53 and DNA damage-inducible expression of the xeroderma pigmentosum group C gene, Proc. Natl. Acad. Sci. U. S. A., 2002, 99, 12985–12990.
M. L. Smith, Y. R. Seo, p53 regulation of DNA excision repair pathways, Mutagenesis, 2002, 17, 149–156.
L. Latonen, M. Laiho, Cellular UV damage responses - functions of tumor suppressor p53, Biochim. Biophys. Acta, 2005, 1755, 71–89.
D. Decraene, P. Agostinis, A. Pupe, P. de Haes, M. Garmyn, Acute response of human skin to solar radiation: regulation and function of the p53 protein, J. Photochem. Photobiol., B, 2001, 63, 78–83 B - Biology.
L. A. de, E. D. Kroon, F. R. de Gruijl, Cell cycle effects and concomitant p53 expression in hairless murine skin after longwave UVA (365 nm) irradiation: a comparison with UVB irradiation, Photochem. Photobiol., 1997, 65, 730–735.
P. M. Girard, M. Pozzebon, F. Delacote, T. Douki, V. Smirnova, E. Sage, Inhibition of S-phase progression triggered by UVA-induced ROS does not require a functional DNA damage checkpoint response in mammalian cells, DNA Repair, 2008, 7, 1500–1516.
G. Maga, G. Villani, E. Crespan, U. Wimmer, E. Ferrari, B. Bertocci, U. Hubscher, 8-oxo-guanine bypass by human DNA polymerases in the presence of auxiliary proteins, Nature, 2007, 447, 606–608.
J. L. Rizzo, J. Dunn, A. Rees, T. M. Rünger, No formation of DNA double-strand breaks and no activation of recombination repair with UVA, J. Invest. Dermatol., 2011, 131, 1139–1148.
T. Douki, A. Reynaud-Angelin, J. Cadet, E. Sage, Bipyrimidine photoproducts rather than oxidative lesions are the main type of DNA damage involved in the genotoxic effect of solar UVA radiation, Biochemistry, 2003, 42, 9221–9226.
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Rünger, T.M., Farahvash, B., Hatvani, Z. et al. Comparison of DNA damage responses following equimutagenic doses of UVA and UVB: A less effective cell cycle arrest with UVA may render UVA-induced pyrimidine dimers more mutagenic than UVB-induced ones. Photochem Photobiol Sci 11, 207–215 (2012). https://doi.org/10.1039/c1pp05232b
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DOI: https://doi.org/10.1039/c1pp05232b