Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult.
OBJECTIVE
Oxidative stress (OS) is believed to be a major contributor to age-related cataract and other age-related diseases.


METHODS
cDNA microarrays were used to identify the spectrum and range of genes with transcript levels that are altered in response to acute H(2)O(2)-induced OS in human lens epithelial (HLE) cells. HLE cells were treated with 50 microM H(2)O(2) for 1 hour in the absence of serum, followed by a return to complete medium. RNAs were prepared from treated and untreated cells at 0, 1, 2, and 8 hours after H(2)O(2) treatment.


RESULTS
The data showed 1171 genes that were significantly up- and downregulated in response to H(2)O(2) treatment. Several functional subcategories of genes were identified, including those encoding DNA repair proteins, antioxidant defense enzymes, molecular chaperones, protein biosynthesis enzymes, and trafficking and degradation proteins. Differential expression of selected genes was confirmed at the level of RNA and/or protein. Many of the identified genes (e.g., glutathione S-transferase [MGST2], thioredoxin reductase beta, and peroxiredoxin 2) have been identified as participants in OS responses in the lens and other systems. Some genes induced by OS in the current study (e.g., oxygen regulated protein [ORP150] and heat shock protein [HSP40]) are better known to respond to other forms of stress. Two genes (receptor tyrosine kinase [AXL/ARK] and protein phosphatase 2A) are known to be differentially expressed in cataract. Most of the genes point to a novel pathways associated with OS.


CONCLUSIONS
The present data provide a global perspective on those genes that respond to acute OS, point to novel genes and pathways associated with OS, and set the groundwork for understanding the functions of OS-related genes in lens protection and disease.
Goswami
Sumanta
S
Department of Ophthalmology and Visual Sciences, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Sheets
Nancy L
NL
Zavadil
Jiri
J
Chauhan
Bharesh K
BK
Bottinger
Erwin P
EP
Reddy
Venkat N
VN
Kantorow
Marc
M
Cvekl
Ales
A
D017209:Apoptosis; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002470:Cell Survival; D003459:Crystallins / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007908:Lens, Crystalline / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress; D012313:RNA / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D003459:Crystallins; D017931:DNA Primers; D012313:RNA; D006861:Hydrogen Peroxide