Proteome analysis of ultraviolet-B-induced protein expression in vitro human dermal fibroblasts

Photodermatol Photoimmunol Photomed. 2010 Dec;26(6):318-26. doi: 10.1111/j.1600-0781.2010.00556.x.

Abstract

Background: Ultraviolet-B (UVB) radiation can result in acute photodamage, photoaging and skin cancer through the induction of reactive oxygen species, DNA damage, activation of signaling pathways, and regulation of gene expression. In this study, we investigated UVB-induced alterations in protein expression in human dermal fibroblasts.

Methods: Skin fibroblasts were irradiated with 100 mJ/cm² UVB, and cell viability was monitored by the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide assay. Two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectroscopy were used to identify differentially expressed proteins. The mRNA and levels of identified proteins were detected using a quantitative real-time polymerase chain reaction assay and Western blot.

Results: UVB decreased the viability of skin fibroblasts. In UVB-treated cells, eighteen differentially expressed proteins were identified. Among these proteins, the amounts of receptor-interacting protein (RIP) and vimentin were significantly up-regulated. However, their mRNA levels decreased and remained relatively stable, respectively.

Conclusions: The differential expression of RIP and vimentin was validated in UVB-irradiated fibroblasts. RIP may promote cell injury, and vimentin may contribute to the resistance of cells to UVB-induced damage.

MeSH terms

  • Adolescent
  • Adult
  • Cell Survival / radiation effects
  • Cells, Cultured
  • Dermis / metabolism*
  • Dermis / pathology
  • Female
  • Fibroblasts / metabolism*
  • Fibroblasts / pathology
  • Gene Expression Regulation / radiation effects*
  • Humans
  • In Vitro Techniques
  • Male
  • Proteome / biosynthesis*
  • Ultraviolet Rays / adverse effects*

Substances

  • Proteome