Microarray analysis of H2O2-, HNE-, or tBH-treated ARPE-19 cells

• Published in: Free radical biology & medicine 2002-11, Vol.33 (10), p.1419-1432
• Main Authors: Weigel, Andrea L, Handa, James T, Hjelmeland, Leonard M
• Format: Article
• Language: English
• Subjects: Apoptosis; Cell Communication; Cell Cycle; Gene Expression Regulation; Humans; Hydrogen Peroxide - pharmacology; Oligonucleotide Array Sequence Analysis; Oxidative Stress; Pigment Epithelium of Eye - cytology; Reverse Transcriptase Polymerase Chain Reaction; RNA - metabolism; Signal Transduction; tert-Butylhydroperoxide - pharmacology; Up-Regulation
• ISSN: 0891-5849
• DOI: 10.1016/s0891-5849(02)01082-1

Oxidative stress plays a key role in aging diseases of the posterior pole of the eye such as age-related macular degeneration. The oxidative stress response of in vitro RPE cells has been studied for a small number of genes. However, a comprehensive transcriptional response has yet to be elucidated. The purpose of this study was to determine if the transcription of a common set of genes is altered by exposure of ARPE-19 cells to three major generators of oxidative stress, hydrogen peroxide (H2O2), 4-hydroxynonenal (HNE), and tert-butylhydroperoxide (tBH). As expected, a common response was observed that included 35 genes differentially regulated by all three treatments. Of these, only one gene was upregulated, and only by one oxidant, while all other responses were downregulation. The majority of these genes fell into five functional categories: apoptosis, cell cycle regulation, cell-cell communication, signal transduction, and transcriptional regulation. Additionally, a large number of genes were differentially regulated by one oxidant only, including the majority of the conventional oxidative stress response genes present on the Clontech Human 1.2 microarray. This study raises questions regarding the generality of results that involve the use of a single oxidant and a single cell culture condition.