Oxidative stress induces differential gene expression in a human lens epithelial cell line

Deborah A. Carper, Jennifer K. Sun, Takeshi Iwata, J. Samuel Zigler, Nobuhiro Ibaraki, Li Ren Lin, Venkat Ready

Research output: Contribution to journalArticle

Abstract

PURPOSE. To identify differentially expressed genes in a human lens epithelial cell line exposed to oxidative stress. METHODS. Reverse transcriptase-polymerase chain reaction (RT-PCR) differential display was used to evaluate differential gene expression in a human lens epithelial cell line (SRA 01-04) when cells were exposed for 3 hours to a single bolus of 200 μM hydrogen peroxide. Differentially expressed genes were identified through DNA sequencing and a nucleotide database search. Differential expression was confirmed by northern blot and RT-PCR analyses. RESULTS. Using 18 primer sets, 28 RT-PCR products were differentially expressed between control and hydrogen peroxide-treated cells. In stressed cells, mitochondrial transcripts nicotinamide adenine dinucleotide (NADH) dehydrogenase subunit 4 and cytochrome b were downregulated 4-fold. Of the cytoplasmic mRNAs, glutamine cyclotransferase decreased 10-fold, whereas cytokine-inducible nuclear protein, alternative splicing factor 2, and β-hydroxyisobutyryl-coenzyme A hydrolase increased 2-, 4-, and 10-fold, respectively. Analysis of mitochondrial transcripts in a 24-hour time course showed that NADH dehydrogenase subunit 4 mRNA decreased by 2-fold as early as 1 hour after oxidative stress, whereas the rate of decrease was slower for cytochrome b, cytochrome oxidase III, and 16S rRNA. CONCLUSIONS. Oxidative stress induced specific expressed gene changes in hydrogen peroxide-treated lens cells, including genes involved in cellular respiration and mRNA and peptide processing. These early changes may reflect pathways involved in the defense, pathology, or both of the lens epithelium, which is exposed to oxidative stress throughout life.

Original languageEnglish (US)
Pages (from-to)400-406
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume40
Issue number2
StatePublished - Feb 1999
Externally publishedYes

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Lenses
Oxidative Stress
Epithelial Cells
Reverse Transcriptase Polymerase Chain Reaction
Hydrogen Peroxide
Gene Expression
Cell Line
Cytochromes b
glutaminyl-peptide cyclotransferase
Messenger RNA
Genes
Protein Splicing
Cell Respiration
Alternative Splicing
Hydrolases
Electron Transport Complex IV
Coenzyme A
Nuclear Proteins
DNA Sequence Analysis
Northern Blotting

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Carper, D. A., Sun, J. K., Iwata, T., Zigler, J. S., Ibaraki, N., Lin, L. R., & Ready, V. (1999). Oxidative stress induces differential gene expression in a human lens epithelial cell line. Investigative Ophthalmology and Visual Science, 40(2), 400-406.

Oxidative stress induces differential gene expression in a human lens epithelial cell line. / Carper, Deborah A.; Sun, Jennifer K.; Iwata, Takeshi; Zigler, J. Samuel; Ibaraki, Nobuhiro; Lin, Li Ren; Ready, Venkat.

In: Investigative Ophthalmology and Visual Science, Vol. 40, No. 2, 02.1999, p. 400-406.

Research output: Contribution to journalArticle

Carper, DA, Sun, JK, Iwata, T, Zigler, JS, Ibaraki, N, Lin, LR & Ready, V 1999, 'Oxidative stress induces differential gene expression in a human lens epithelial cell line', Investigative Ophthalmology and Visual Science, vol. 40, no. 2, pp. 400-406.
Carper DA, Sun JK, Iwata T, Zigler JS, Ibaraki N, Lin LR et al. Oxidative stress induces differential gene expression in a human lens epithelial cell line. Investigative Ophthalmology and Visual Science. 1999 Feb;40(2):400-406.
Carper, Deborah A. ; Sun, Jennifer K. ; Iwata, Takeshi ; Zigler, J. Samuel ; Ibaraki, Nobuhiro ; Lin, Li Ren ; Ready, Venkat. / Oxidative stress induces differential gene expression in a human lens epithelial cell line. In: Investigative Ophthalmology and Visual Science. 1999 ; Vol. 40, No. 2. pp. 400-406.
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