title: Proteomic analysis of UVB-induced protein expression- and redox-dependent changes in skin fibroblasts using lysine- and cysteine-labeling two-dimensional difference gel electrophoresis.
abstract: UVB is the most energetic and DNA-damaging to humans in ultraviolet radiation. Previous research has suggested that exposure to UVB causes skin pathologies because of direct DNA damage and the generation of reactive oxygen species (ROS). However, the detailed molecular mechanisms by which UVB leads to skin cancer have yet to be clarified. In the current study, normal skin fibroblast cells (CCD-966SK) were exposed to various doses of UVB, and the changes in protein expression and thiol reactivity were monitored with lysine- and cysteine-labeling 2D-DIGE and MALDI-TOF mass spectrometry. Our proteomic analysis revealed that 89 identified proteins showed significant changes in protein expression, and 37 in thiol reactivity. Many proteins that are known to be involved in protein folding, redox regulation and nucleotide biosynthesis were up-regulated under UVB irradiation. In contrast, proteins responsible for biosynthesis and protein degradation were down-regulated. In addition, the thiol-reactivity of proteins involving cytoskeleton, metabolism, and signal transduction were altered by UVB. In summary, these UVB-modulated cellular proteins and redox-regulated proteins might play important roles in the early stages of skin cancer formation and photoaging induced by UVB-irradiation. Such proteins might provide a potential target for the rational design of drugs to prevent UVB-induced diseases.
authors.affiliation: Institute of Bioinformatics and Structural Biology & Department of Medical Sciences, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chieh-Lin
authors.initials: CL
authors.lastname: Wu
authors.forename: Hsiu-Chuan
authors.initials: HC
authors.lastname: Chou
authors.forename: Chao-Sheng
authors.initials: CS
authors.lastname: Cheng
authors.forename: Ji-Min
authors.initials: JM
authors.lastname: Li
authors.forename: Szu-Ting
authors.initials: ST
authors.lastname: Lin
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Chen
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Chan
mesh_terms: D002460:Cell Line; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000528:radiation effects*; D006801:Humans; D059748:Proteolysis / Q000528:radiation effects*; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012867:Skin / Q000378:metabolism*; D012878:Skin Neoplasms / Q000378:metabolism; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods; D014466:Ultraviolet Rays
chemical_list: D017382:Reactive Oxygen Species