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S. Matsuda, F. Gomi, T. Katayama, M. Tohyama, Y. Tano; Identification of Genes Expressed by Oxidative Stress in Retinal Pigment Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1634.
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Purpose: To isolate the genes whose expression increases in retinal pigment epithelial (RPE) cells due to oxidative stress. Methods: Subconfluent cultures of ARPE-19 cells were treated with hypoxia/re-oxygenation or paraquat to induce oxidative stress. The mRNA was isolated from cells, and genes were screened whose expression increased due to oxidative stress using a cDNA subtraction method. The change of expression of candidate genes was confirmed by Northern blotting or reverse transcriptase-polymerase chain reaction (RT-PCR). The expression of several genes was also analyzed in mice treated with ocular focal illumination after intraperitoneal injection of Rose Bengal photosensitive dye as a model for ocular responses to oxidative stress by RT-PCR. Results: The up-regulation of 10 genes in response to oxidative stress was confirmed and included connective tissue growth factor (CTGF) and matrix metalloproteinase-2 (MMP-2). The peak expression of MMP-2 transcripts occurred at an earlier period after treatment with oxidative stress compared with CTGF. The induction of CTGF and MMP2 transcripts also occurred in the eyes of mice treated with Rose Bengal and light. Conclusions: The expression of CTGF and MMP-2 mRNA increased in the RPE cells and in vivo after treatment with oxidative stress. Further analysis may provide a better understanding of the pathogenesis of age-related macular degeneration.
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