Proteomic analysis of UVB-induced protein expression- and redox-dependent changes in skin fibroblasts using lysine- and cysteine-labeling two-dimensional difference gel electrophoresis

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.

Graphical abstractFigure optionsDownload full-size imageDownload high-quality image (182 K)Download as PowerPoint slideHighlights► UVB-induced changes in protein expression and thiol reactivity were monitored with lysine- and cysteine-labeling 2D-DIGE. ► In this study, 89 identified proteins showed significant changes in protein expression, and 37 in thiol reactivity. ► Proteins involved in folding, redox regulation, and degradation were altered in expression under UVB irradiation. ► Proteins involved in cytoskeleton, metabolism, and signaling were altered in thiol reactivity under UVB irradiation. ► UVB plays important roles in the early stages of skin cancer formation and photoaging.