Density-dependent expression of FGF-2 in response to oxidative stress in RPE cells in vitro
| Autor: | Mitsumasa Wada, Claire M. Gelfman, Mitra Alizadeh, James T. Handa, Leonard M. Hjelmeland, Lawrence S Morse, Hiroshi Matsunaga |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Cell Survival
Basic fibroblast growth factor Cell Count Biology medicine.disease_cause Cellular and Molecular Neuroscience chemistry.chemical_compound tert-Butylhydroperoxide medicine Humans RNA Messenger Viability assay Pigment Epithelium of Eye Fibroblast Cells Cultured chemistry.chemical_classification Reactive oxygen species Dose-Response Relationship Drug Membrane Proteins Hydrogen Peroxide Blotting Northern Molecular biology Sensory Systems Cell biology Blot Oxidative Stress Ophthalmology medicine.anatomical_structure chemistry Cell culture Heme Oxygenase (Decyclizing) Fibroblast Growth Factor 2 Reactive Oxygen Species Heme Oxygenase-1 Intracellular Oxidative stress |
| Zdroj: | Current Eye Research. 23:226-231 |
| ISSN: | 1460-2202 0271-3683 |
| Popis: | The purpose of this study is to demonstrate the effect of culture density on the steady state mRNA levels of fibroblast growth factor-2 (FGF-2) when retinal pigment epithelial (RPE) cells are subjected to oxidative stress in vitro.Subconfluent and confluent cultures of the established RPE cell line ARPE-19, were treated with increasing concentrations of tert-butyl hydroperoxide (tBH) or hydrogen peroxide (H(2)O(2)). Cell viability was measured using the WST-1 assay, and intracellular reactive oxygen intermediate (ROI) production was quantified by dichlorofluoroscein (DCF) fluorescence. Steady state changes in heme oxygenase-1 (HO-1) and FGF-2 mRNAs were measured by Northern blot analysis.Confluent cultures of ARPE-19 cells were less susceptible than subconfluent cultures to the toxic effects of the chemical oxidants. The intracellular reactive oxygen intermediate production was higher in subconfluent than confluent cultures with increasing tBH concentration. At nontoxic concentrations of tBH and H(2)O(2), a dose dependent increase in FGF-2 expression was seen as a function of culture density. FGF-2 mRNA expression was induced after tBH treatment in subconfluent, but not confluent cells. On the other hand, FGF-2 mRNA induction was observed after H(2)O( 2) treatment in confluent, but not subconfluent cultures. In contrast, no density dependent induction of HO-1 mRNA was seen after treatment with either tBH or H(2)O(2).These results suggest that care should be taken to control for cell density in similar types of in vitro experiments. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|