Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult

Sumanta Goswami, Nancy L. Sheets, Jiři Zavadil, Bharesh K. Chauhan, Erwin P. Bottinger, Venkat N. Reddy, Marc Kantorow, Ales Cvekl

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

PURPOSE. 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 H2O2-induced OS in human lens epithelial (HLE) cells. HLE cells were treated with 50 μM H2O2 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 H2O2 treatment. RESULTS. The data showed 1171 genes that were significantly up- and downregulated in response to H2O2 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 β, 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.

Original languageEnglish (US)
Pages (from-to)2084-2093
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume44
Issue number5
DOIs
StatePublished - May 1 2003

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Lenses
Oxidative Stress
Epithelial Cells
Genes
Cataract
Thioredoxin Reductase 2
Lens Diseases
HSP40 Heat-Shock Proteins
RNA
Peroxiredoxins
Protein Phosphatase 2
Molecular Chaperones
Receptor Protein-Tyrosine Kinases
Protein Biosynthesis
Protein Transport
Enzymes
Oligonucleotide Array Sequence Analysis
Glutathione Transferase
DNA Repair
Proteolysis

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult. / Goswami, Sumanta; Sheets, Nancy L.; Zavadil, Jiři; Chauhan, Bharesh K.; Bottinger, Erwin P.; Reddy, Venkat N.; Kantorow, Marc; Cvekl, Ales.

In: Investigative Ophthalmology and Visual Science, Vol. 44, No. 5, 01.05.2003, p. 2084-2093.

Research output: Contribution to journalArticle

Goswami, S, Sheets, NL, Zavadil, J, Chauhan, BK, Bottinger, EP, Reddy, VN, Kantorow, M & Cvekl, A 2003, 'Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult', Investigative Ophthalmology and Visual Science, vol. 44, no. 5, pp. 2084-2093. https://doi.org/10.1167/iovs.02-0882
Goswami, Sumanta ; Sheets, Nancy L. ; Zavadil, Jiři ; Chauhan, Bharesh K. ; Bottinger, Erwin P. ; Reddy, Venkat N. ; Kantorow, Marc ; Cvekl, Ales. / Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult. In: Investigative Ophthalmology and Visual Science. 2003 ; Vol. 44, No. 5. pp. 2084-2093.
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AU - Goswami, Sumanta

AU - Sheets, Nancy L.

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AU - Chauhan, Bharesh K.

AU - Bottinger, Erwin P.

AU - Reddy, Venkat N.

AU - Kantorow, Marc

AU - Cvekl, Ales

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N2 - PURPOSE. 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 H2O2-induced OS in human lens epithelial (HLE) cells. HLE cells were treated with 50 μM H2O2 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 H2O2 treatment. RESULTS. The data showed 1171 genes that were significantly up- and downregulated in response to H2O2 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 β, 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.

AB - PURPOSE. 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 H2O2-induced OS in human lens epithelial (HLE) cells. HLE cells were treated with 50 μM H2O2 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 H2O2 treatment. RESULTS. The data showed 1171 genes that were significantly up- and downregulated in response to H2O2 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 β, 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.

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