Photogenotoxicity of Mammalian Cells: A Review of the Different Assays for In Vitro Testing¶

Abstract During the past several years, phototoxicity has been studied at the molecular level, and these studies have provided new insights in the field of DNA lesion characterization, DNA repair and cell response to ultraviolet (UV)-induced stress. The development of new antibiotics and antiinflammatory drugs has highlighted the necessity to develop the assessment of phototoxicity in the safety evaluation of new chemical compounds. This paper aims at reviewing the known molecular mechanisms of the cellular response to UV-induced stress, the in vitro methods that can be proposed and used to screen for toxicity of sunlight and the photosensitization process resulting from the activation of drugs by light. UV sources, biological systems and endpoints of interest in that particular objective are listed. Phototoxic effects span from the cytotoxic–apoptotic effect to the induction of primary DNA damage, DNA repair and a variety of stress genes acting on the cell cycle and the fate of the cell. Ultimately, it can lead to the induction of hereditary DNA modification. A variety of assays are proposed to specifically address all these particular consequences of UV-induced toxicity.

[1]  A. Fourtanier,et al.  UVAI-induced Edema and Pyrimidine Dimers in Murine Skin¶ , 2000, Photochemistry and photobiology.

[2]  N. Traynor,et al.  Photogenotoxicity of Skin Phototumorigenic Fluoroquinolone Antibiotics Detected Using the Comet Assay , 1997, Photochemistry and photobiology.

[3]  F. Bernerd,et al.  Evaluation of the Protective Effect of Sunscreens on In Vitro Reconstructed Human Skin Exposed to UVB or UVA Irradiation , 2000, Photochemistry and photobiology.

[4]  J. Cadet,et al.  Use of the single-cell gel electrophoresis assay for the immunofluorescent detection of specific DNA damage. , 1998, Analytical biochemistry.

[5]  A. Sancar,et al.  DNA repair enzymes. , 1988, Annual review of biochemistry.

[6]  D. Schmitt,et al.  Integration of Langerhans cells into a pigmented reconstructed human epidermis. , 1997, The Journal of investigative dermatology.

[7]  L. Marrot,et al.  Differences in the photogenotoxic potential of two fluoroquinolones as shown in diploid yeast strain (Saccharomyces cerevisae) and supercoiled plasmid DNA. , 2000, Mutation research.

[8]  L. Marrot,et al.  The Human Melanocyte as a Particular Target for UVA Radiation and an Endpoint for Photoprotection Assessment , 1999, Photochemistry and photobiology.

[9]  M Liebsch,et al.  The International EU/COLIPA In Vitro Phototoxicity Validation Study: Results of Phase II (Blind Trial). Part 1: The 3T3 NRU Phototoxicity Test. , 1998, Toxicology in vitro : an international journal published in association with BIBRA.

[10]  F. Urbach,et al.  Ultraviolet radiation and skin cancer of humans. , 1997, Journal of photochemistry and photobiology. B, Biology.

[11]  K J Johanson,et al.  Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. , 1984, Biochemical and biophysical research communications.

[12]  C. Potten,et al.  The detection of cyclobutane thymine dimers, (6-4) photolesions and the Dewar photoisomers in sections of UV-irradiated human skin using specific antibodies, and the demonstration of depth penetration effects. , 1995, Journal of photochemistry and photobiology. B, Biology.

[13]  J. Cadet,et al.  Formation of the Main UV-induced Thymine Dimeric Lesions within Isolated and Cellular DNA as Measured by High Performance Liquid Chromatography-Tandem Mass Spectrometry* , 2000, The Journal of Biological Chemistry.

[14]  G. Núñez,et al.  Caspases: the proteases of the apoptotic pathway , 1998, Oncogene.

[15]  D. Lane,et al.  High levels of p53 protein in UV-irradiated normal human skin. , 1993, Oncogene.

[16]  P. Kasper,et al.  The application of the micronucleus test in Chinese hamster V79 cells to detect drug-induced photogenotoxicity. , 1999, Mutation research.

[17]  P. Schmezer,et al.  The effect of various antioxidants and other modifying agents on oxygen-radical-generated DNA damage in human lymphocytes in the COMET assay. , 1994, Mutation research.

[18]  N. Dumaz,et al.  p53 mutations in skin and internal tumors of xeroderma pigmentosum patients belonging to the complementation group C. , 1998, Cancer research.

[19]  B. Epe,et al.  Wavelength dependence of oxidative DNA damage induced by UV and visible light. , 1997, Carcinogenesis.

[20]  E. Drobetsky,et al.  A role for ultraviolet A in solar mutagenesis. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  L. Klotz,et al.  Evidence that Singlet Oxygen-induced Human T Helper Cell Apoptosis Is the Basic Mechanism of Ultraviolet-A Radiation Phototherapy , 1997, The Journal of experimental medicine.

[22]  E. Sage,et al.  Use of the alkaline comet assay to detect DNA repair deficiencies in human fibroblasts exposed to UVC, UVB, UVA and gamma-rays. , 1996, International journal of radiation biology.

[23]  J. Cooper,et al.  Quantification of active caspase 3 in apoptotic cells. , 2000, Analytical biochemistry.

[24]  P. Hanawalt Transcription-coupled repair and human disease. , 1994, Science.

[25]  L. Mullenders,et al.  Transcription‐coupled repair removes both cyclobutane pyrimidine dimers and 6‐4 photoproducts with equal efficiency and in a sequential way from transcribed DNA in xeroderma pigmentosum group C fibroblasts. , 1995, The EMBO journal.

[26]  J. Cadet,et al.  Oxidative damage to DNA: formation, measurement, and biological significance. , 1997, Reviews of physiology, biochemistry and pharmacology.

[27]  P. Burcham Genotoxic lipid peroxidation products: their DNA damaging properties and role in formation of endogenous DNA adducts. , 1998, Mutagenesis.

[28]  N. Davidson,et al.  Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis. , 1993, Cancer research.

[29]  N. Traynor,et al.  The phototumorigenic fluoroquinolone, lomefloxacin, photosensitises p53 accumulation and transcriptional activity in human skin cells. , 2000, Journal of photochemistry and photobiology. B, Biology.

[30]  D. Green,et al.  Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl , 1995, The Journal of experimental medicine.

[31]  B. Carnes,et al.  INDUCTION OF DIRECT AND INDIRECT SINGLE‐STRAND BREAKS IN HUMAN CELL DNA BY FAR‐ AND NEAR‐ULTRAVIOLET RADIATIONS: ACTION SPECTRUM AND MECHANISMS , 1987, Photochemistry and photobiology.

[32]  R E Durand,et al.  Factors influencing DNA migration from individual cells subjected to gel electrophoresis. , 1992, Experimental cell research.

[33]  D. Lane,et al.  p53, guardian of the genome , 1992, Nature.

[34]  J. Hoeijmakers,et al.  Different removal of ultraviolet photoproducts in genetically related xeroderma pigmentosum and trichothiodystrophy diseases. , 1995, Cancer research.

[35]  J. Simon,et al.  A role for sunlight in skin cancer: UV-induced p53 mutations in squamous cell carcinoma. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[36]  A. Sarasin,et al.  Clues to epidermal cancer proneness revealed by reconstruction of DNA repair-deficient xeroderma pigmentosum skin in vitro , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[37]  C. Provot,et al.  In vitro eukaryotic DNA excision repair assays: an overview. , 1995, Biochimie.

[38]  D. Averbeck Photoaddition by Furocoumarins , 1994 .

[39]  S Albertini,et al.  The photomutagenicity of fluoroquinolones and other drugs. , 1998, Toxicology letters.

[40]  D. Green,et al.  The Release of Cytochrome c from Mitochondria: A Primary Site for Bcl-2 Regulation of Apoptosis , 1997, Science.

[41]  F. Bernerd,et al.  UVA exposure of human skin reconstructed in vitro induces apoptosis of dermal fibroblasts: subsequent connective tissue repair and implications in photoaging , 1998, Cell Death and Differentiation.

[42]  D. Averbeck,et al.  Genetic effect of 3-carbethoxypsoralen, angelicin, psoralen and 8-methoxypsoralen plus 365-nm irradiation in Saccharomyces cerevisiae: induction of reversions, mitotic crossing-over, gene conversion and cytoplasmic "petite" mutations. , 1979, Mutation research.

[43]  S. Srinivasula,et al.  Cytochrome c and dATP-Dependent Formation of Apaf-1/Caspase-9 Complex Initiates an Apoptotic Protease Cascade , 1997, Cell.

[44]  Xiaodong Wang,et al.  Apaf-1, a Human Protein Homologous to C. elegans CED-4, Participates in Cytochrome c–Dependent Activation of Caspase-3 , 1997, Cell.

[45]  M. Fenech,et al.  The cytokinesis-block micronucleus technique: a detailed description of the method and its application to genotoxicity studies in human populations. , 1993, Mutation research.

[46]  R. Tice,et al.  A simple technique for quantitation of low levels of DNA damage in individual cells. , 1988, Experimental cell research.

[47]  M. Nüsse,et al.  UV-B-induced cell cycle perturbations, micronucleus induction, and modulation by caffeine in human keratinocytes. , 1996, International journal of radiation biology.

[48]  R. Snyder,et al.  Considerations on photochemical genotoxicity: Report of the International Workshop on Genotoxicity Test Procedures Working Group , 2000, Environmental and molecular mutagenesis.

[49]  E. Sage DISTRIBUTION AND REPAIR OF PHOTOLESIONS IN DNA: GENETIC CONSEQUENCES AND THE ROLE OF SEQUENCE CONTEXT , 1993, Photochemistry and photobiology.

[50]  B. Epe,et al.  Oxidative DNA damage induced by visible light in mammalian cells: extent, inhibition by antioxidants and genotoxic effects. , 1998, Mutation research.

[51]  D. Harrison,et al.  Apoptosis: an overview of the process and its relevance in disease. , 1997, Advances in pharmacology.

[52]  Horst Spielmann,et al.  In Vitro Phototoxicity Testing , 1994 .

[53]  M. Kirsch‐Volders,et al.  The in vitro micronucleus test: a multi-endpoint assay to detect simultaneously mitotic delay, apoptosis, chromosome breakage, chromosome loss and non-disjunction. , 1997, Mutation research.

[54]  D. Green,et al.  Caspase-3 Is the Primary Activator of Apoptotic DNA Fragmentation via DNA Fragmentation Factor-45/Inhibitor of Caspase-activated DNase Inactivation* , 1999, The Journal of Biological Chemistry.

[55]  R. Tyrrell,et al.  Induction of oxidative DNA base damage in human skin cells by UV and near visible radiation. , 1997, Carcinogenesis.

[56]  W. Maltzman,et al.  UV irradiation stimulates levels of p53 cellular tumor antigen in nontransformed mouse cells , 1984, Molecular and cellular biology.

[57]  L. Cerroni,et al.  p53 mutation in squamous cell carcinomas from psoriasis patients treated with psoralen + UVA (PUVA). , 1997, The Journal of investigative dermatology.

[58]  N. Dumaz,et al.  Quantitative detection of ultraviolet-specific p53 mutations in normal skin from Japanese patients. , 1997, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[59]  B. Epe,et al.  Visible light generates oxidative DNA base modifications in high excess of strand breaks in mammalian cells. , 1994, Carcinogenesis.

[60]  L. Marrot,et al.  An in vitro strategy to evaluate the phototoxicity of solar UV at the molecular and cellular level: application to photoprotection assessment. , 1998, European journal of dermatology : EJD.

[61]  D. Averbeck,et al.  The Second ECVAM Workshop on Phototoxicity Testing , 2000, Alternatives to laboratory animals : ATLA.

[62]  S. Queillé,et al.  Effects of XPD mutations on ultraviolet-induced apoptosis in relation to skin cancer-proneness in repair-deficient syndromes. , 2001, The Journal of investigative dermatology.

[63]  S. Queillé,et al.  High levels of patched gene mutations in basal-cell carcinomas from patients with xeroderma pigmentosum. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[64]  J. Cadet,et al.  Measurement of oxidative damage at pyrimidine bases in gamma-irradiated DNA. , 1996, Chemical research in toxicology.

[65]  A. Sarasin,et al.  The molecular pathways of ultraviolet-induced carcinogenesis. , 1999, Mutation research.

[66]  Xiaodong Wang,et al.  Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c , 1996, Cell.

[67]  Michael Balls,et al.  A Study on UV Filter Chemicals from Annex VII of European Union Directive 76/768/EEC, in the In Vitro 3T3 NRU Phototoxicity Test , 1998, Alternatives to laboratory animals : ATLA.

[68]  M. Green,et al.  Induction of cyclobutane pyrimidine dimers, pyrimidine(6-4)pyrimidone photoproducts, and Dewar valence isomers by natural sunlight in normal human mononuclear cells. , 1995, Cancer research.

[69]  K. Herbert,et al.  Immunochemical quantitation of UV-induced oxidative and dimeric DNA damage to human keratinocytes , 2000, Free radical research.

[70]  A. Collins,et al.  Oxidative damage to DNA: do we have a reliable biomarker? , 1996, Environmental health perspectives.

[71]  P. Kasper,et al.  The Relevance of Photomutagenicity Testing as a Predictor of Photocarcinogenicity , 1998 .

[72]  J. Radicella,et al.  Characterization of the hOGG1 promoter and its expression during the cell cycle. , 2000, Mutation research.

[73]  R. Tice,et al.  Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing , 2000, Environmental and molecular mutagenesis.

[74]  G. Emri,et al.  Low doses of UVB or UVA induce chromosomal aberrations in cultured human skin cells. , 2000, The Journal of investigative dermatology.

[75]  N. Dumaz,et al.  The specificity of p53 mutation spectra in sunlight induced human cancers. , 1995, Journal of photochemistry and photobiology. B, Biology.

[76]  A. Sarasin,et al.  RETRACTED: Transcription-Coupled Repair of 8-oxoGuanine Requirement for XPG, TFIIH, and CSB and Implications for Cockayne Syndrome , 2000, Cell.

[77]  E. Sage,et al.  Wavelength dependence of ultraviolet-induced DNA damage distribution: involvement of direct or indirect mechanisms and possible artefacts. , 1999, Journal of photochemistry and photobiology. B, Biology.

[78]  A. Sarasin,et al.  Cell survival and shuttle vector mutagenesis induced by ultraviolet A and ultraviolet B radiation in a human cell line. , 1996, The Journal of investigative dermatology.

[79]  A. Sarasin,et al.  Variability in nucleotide excision repair and cancer risk: a review. , 2000, Mutation research.