title: DCT protects human melanocytic cells from UVR and ROS damage and increases cell viability.
abstract: Dopachrome tautomerase (DCT) is involved in the formation of the photoprotective skin pigment eumelanin and has also been shown to have a role in response to apoptotic stimuli and oxidative stress. The effect of DCT on UVR DNA damage responses and survival pathways in human melanocytic cells was examined by knockdown experiments using melanoma cells, neonatal foreskin melanoblasts (MB) in monoculture and in co-culture with human keratinocytes. MB cell strains genotyped as either MC1R WT or MC1R RHC homozygotes, which are known to be deficient in DCT, were transduced with lentivirus vectors for either DCT knockdown or overexpression. We found melanoma cell survival was reduced by DCT depletion and by UVR over time. UVR-induced p53 and pp53-Ser15 levels were reduced with DCT depletion. Knockdown of DCT in MC1R WT and MC1R RHC MB cells reduced their survival after UVR exposure, whereas increased DCT protein levels enhanced survival. DCT depletion reduced p53 and pp53-Ser15 levels in WM266-4 melanoma and MC1R WT MB cells, while MC1R RHC MB cells displayed variable levels. Both MC1R WT and RHC genotypes of MB cells were responsive to UVR at 3 h with increases in both p53 and pp53-Ser15 proteins. MC1R WT MB cell strains in coculture with keratinocytes have an increased cell survival after UVR exposure when compared to those in monoculture, a protective effect which appears to be conferred by the keratinocytes.
authors.affiliation: Institute for Molecular Bioscience, Melanogenix Group, The University of Queensland, Brisbane, Qld, Australia.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Ainger
authors.forename: Xuan L
authors.initials: XL
authors.lastname: Yong
authors.forename: Shu S
authors.initials: SS
authors.lastname: Wong
authors.forename: Dubravka
authors.initials: D
authors.lastname: Skalamera
authors.forename: Brian
authors.initials: B
authors.lastname: Gabrielli
authors.forename: J Helen
authors.initials: JH
authors.lastname: Leonard
authors.forename: Richard A
authors.initials: RA
authors.lastname: Sturm
mesh_terms: D002470:Cell Survival / Q000528:radiation effects*; D018920:Coculture Techniques; D004249:DNA Damage; D055785:Gene Knockdown Techniques; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism; D008544:Melanocytes / Q000166:cytology / Q000378:metabolism* / Q000528:radiation effects*; D008545:Melanoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D034741:RNA, Small Interfering / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D044102:Receptor, Melanocortin, Type 1 / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays / Q000009:adverse effects*; D015854:Up-Regulation
chemical_list: D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D044102:Receptor, Melanocortin, Type 1; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase
keywords: Dopachrome tautomerase; melanocortin-1 receptor; melanocyte; melanoma; p53; pigmentation