Differential Expression Of Proteins In Human Fibroblasts Induced By Ultraviolet Irradiation

Backgroud: Ultraviolet plays an important role in acute damage, inflammation and photoaging of the skin, which characterized by wrikles, loss of skin tone and resilience, and skin cancer. Based on the wavelength, ultraviolet is divided into three categories (UVA 320-400nm, UVB 280-320nm, UVC 200-280nm) . It has been well documented that ultraviolet irradiation can affect the expression of many proteins in human skin cells. Fibroblasts are important cells in skin structure and physical function. Our lab has discovered 33 differential protein spots between the ultraviolet exposed and unexposed fibroblasts by two-dimensional gel electrophoresis and proteinomic techniques. Among them, several candidate proteins were identified by MALDI-TOF-MS, including up-regulated Vimentin, Death receptor interacting protein, IL-1 receptor accessory protein (IL-1 RAcP) , Prohibitin and Prolyl 4-hydroxylase (PHD4); down-regulated Annexin A2 and Phosphoglycerate kinase (PGK1) . Up-regulated Vimentin and RIP in fibroblasts were further analysed by Western-blotting and immuno-fluorescences in our previous studies.Purpose: To investigate the differential expression of Annexin A2 and PGK.1 mRNA in human fibroblasts induced by ultraviolet. To identify the early markers of ultraviolet induced damage of the skin.Method: Human fibroblasts were isolated from fresh foreskin of health man, and cultured in vitro. Cultured fibroblasts were irradiated by UVB at a dose of 100mJ/cm~2. Total RNA was purified and amplified by RT-PCR after 48h incubation. Then used ultraviolet spectrophotometer for photograph and analyzed the band. Compared the ANX2 and PGK1 to beta actin mRNA for half quantitative ananlysis by SPSS.Result: UVB can down-regulate PGK1 mRNA expression of fibroblast.Conclusion: PGK1 mRNA could be served as the marker of ultraviolet early injury.