Density-dependent expression of FGF-2 in response to oxidative stress in RPE cells in vitro

• DOI: 10.1076/ceyr.23.3.226.5467
• Publication Date: 2003-02-13T14:12:54Z
• Authors: Mitsumasa Wada; Claire Mazow Gelfman; Hiroshi Matsunaga; Mitra Alizadeh; Lawrence Morse; James T. Handa; Leonard M. Hjelme...
• Keywords: Dose-Response Relationship, Drug; Cell Survival; Membrane Proteins; Cell Count; Hydrogen Peroxide; Blotting, Northern; Oxidative Stress; 03 medical and health sciences; 0302 clinical medicine; tert-Butylhydroperoxide; Heme Oxygenase (Decyclizing); Humans; Fibroblast Growth Factor 2; RNA, Messenger; Pigment Epithelium of Eye; Reactive Oxygen Species; Cells, Cultured; Heme Oxygenase-1

AbstractPurpose. The purpose of this study is to demonstrate the effect culture density on steady state mRNA levels fibroblast growth factor-2 (FGF-2) when retinal pigment epithelial (RPE) cells are subjected oxidative stress in vitro. Methods. Subconfluent and confluent cultures established RPE cell line ARPE-19, were treated with increasing concentrations tert-butyl hydroperoxide (tBH) or hydrogen peroxide (H 2 O 2) . Cell viability was measured using WST-1 assay, intracellular reactive oxygen intermediate (ROI) production quantified by dichlorofluoroscein (DCF) fluorescence. Steady changes heme oxygenase-1 (HO-1) FGF-2 mRNAs Northern blot analysis. Results. Confluent ARPE-19 less susceptible than subconfluent toxic effects chemical oxidants. higher cultres tBH concentration. At nontoxic H 2, a dose dependent increase expression seen as function density. induced after treatment subconfluent, but not cells. On other hand, induction observed confluent, cultures. In contrast, no HO-1 either 2. Conclusions. These results suggest that care should be taken control for similar types vitro experiments.