title: Investigating the role of melanin in UVA/UVB- and hydrogen peroxide-induced cellular and mitochondrial ROS production and mitochondrial DNA damage in human melanoma cells.
abstract: Skin cancer incidence is dramatically increasing worldwide, with exposure to ultraviolet radiation (UVR) a predominant factor. The UVA component initiates oxidative stress in human skin, although its exact role in the initiation of skin cancer, particularly malignant melanoma, remains unclear and is controversial because there is evidence for a melanin-dependent mechanism in UVA-linked melanoma studies. Nonpigmented (CHL-1, A375), moderately pigmented (FM55, SKmel23), and highly pigmented (FM94, hyperpigmented FM55) human melanoma cell lines have been used to investigate UVA-induced production of reactive oxygen species using FACS analysis, at both the cellular (dihydrorhodamine-123) and the mitochondrial (MitoSOX) level, where most cellular stress is generated. For the first time, downstream mtDNA damage (utilizing a quantitative long-PCR assay) has been investigated. Using UVA, UVB, and H(2)O(2) as cellular stressors, we have explored the dual roles of melanin as a photoprotector and photosensitizer. The presence of melanin has no influence over cellular oxidative stress generation, whereas, in contrast, melanin protects against mitochondrial superoxide generation and mtDNA damage (one-way ANOVA with post hoc Tukey's analysis, P<0.001). We show that if melanin binds directly to DNA, it acts as a direct photosensitizer of mtDNA damage during UVA irradiation (P<0.001), providing evidence for the dual roles of melanin.
authors.affiliation: Department of Dermatological Sciences, Institute of Cellular Medicine, Medical School, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.
authors.forename: Helen
authors.initials: H
authors.lastname: Swalwell
authors.affiliation: Department of Dermatological Sciences, Institute of Cellular Medicine, Medical School, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Latimer
authors.affiliation: RAFT Leopold Muller Building, Mount Vernon Hospital, Northwood, Middlesex HA6 2RN, UK.
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Haywood
authors.affiliation: Department of Dermatological Sciences, Institute of Cellular Medicine, Medical School, Newcastle University, Newcastle upon Tyne NE2 4HH, UK. Electronic address: m.a.birch-machin@ncl.ac.uk.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Birch-Machin
mesh_terms: D045744:Cell Line, Tumor; D019610:Cytoprotection; D004249:DNA Damage; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008543:Melanins / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D008545:Melanoma; D008928:Mitochondria / Q000378:metabolism* / Q000528:radiation effects; D009281:Naphthalenes; D010082:Oxepins; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014466:Ultraviolet Rays
chemical_list: C461941:1-phenyl-1,4-epoxy-1H,4H-naphtho(1,8-de)(1,2)dioxepin; D004272:DNA, Mitochondrial; D008543:Melanins; D009281:Naphthalenes; D010082:Oxepins; D017382:Reactive Oxygen Species