Quantitative AP-1 gene regulation by oxidative stress in the human retinal pigment epithelium

Edward Chaum, Jinggang Yin, Huaitao Yang, Fridtjof Thomas, John C. Lang

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The purpose of this study was to characterize the early molecular responses to quantified levels of oxidative stress (OS) in the human retinal pigment epithelium (RPE). Confluent ARPE-19 cells were cultured for 3 days in defined medium to stabilize gene expression. The cells were exposed to varying levels of OS (0-500 μMH2O2) for 1-8 h and gene expression was followed for up to 24-h after OS. Using real-time qPCR, we quantified the expression of immediate early genes from the AP-1 transcription factor family and other genes involved in regulating the redox status of the cells. Significant and quantitative changes were seen in the expression of six AP-1 transcription factor genes, FosB, c-Fos, Fra-1, c-Jun, JunB, and ATF3 from 1-8 h following OS. The peak level of induced transcription from OS varied from 2- to 128-fold over the first 4 h, depending on the gene and magnitude of OS. Increased transcription at higher levels of OS was also seen for up to 8-h for some of these genes. Protein translation was examined for 24-h following OS using Western blotting methods, and compared to the qPCR responses. We identified six AP-1 family genes that demonstrate quantitative upregulation of expression in response to OS. Two distinct types of quantifiable OS-specific responses were observed; dose-dependent responses, and threshold responses. Our studies show that different levels of OS can regulate the expression of AP-1 transcription factors quantitatively in the human RPE in vitro.

Original languageEnglish (US)
Pages (from-to)1280-1291
Number of pages12
JournalJournal of Cellular Biochemistry
Volume108
Issue number6
DOIs
StatePublished - Dec 15 2009

Fingerprint

Oxidative stress
Retinal Pigments
Retinal Pigment Epithelium
Transcription Factor AP-1
Gene expression
Oxidative Stress
Genes
Transcription
fos Genes
Gene Expression
Immediate-Early Genes
Protein Biosynthesis
Oxidation-Reduction
Cultured Cells
Up-Regulation
Western Blotting

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Quantitative AP-1 gene regulation by oxidative stress in the human retinal pigment epithelium. / Chaum, Edward; Yin, Jinggang; Yang, Huaitao; Thomas, Fridtjof; Lang, John C.

In: Journal of Cellular Biochemistry, Vol. 108, No. 6, 15.12.2009, p. 1280-1291.

Research output: Contribution to journalArticle

Chaum, Edward ; Yin, Jinggang ; Yang, Huaitao ; Thomas, Fridtjof ; Lang, John C. / Quantitative AP-1 gene regulation by oxidative stress in the human retinal pigment epithelium. In: Journal of Cellular Biochemistry. 2009 ; Vol. 108, No. 6. pp. 1280-1291.
@article{9e3a7c54f28a4747b29bf9f2c1620e67,
title = "Quantitative AP-1 gene regulation by oxidative stress in the human retinal pigment epithelium",
abstract = "The purpose of this study was to characterize the early molecular responses to quantified levels of oxidative stress (OS) in the human retinal pigment epithelium (RPE). Confluent ARPE-19 cells were cultured for 3 days in defined medium to stabilize gene expression. The cells were exposed to varying levels of OS (0-500 μMH2O2) for 1-8 h and gene expression was followed for up to 24-h after OS. Using real-time qPCR, we quantified the expression of immediate early genes from the AP-1 transcription factor family and other genes involved in regulating the redox status of the cells. Significant and quantitative changes were seen in the expression of six AP-1 transcription factor genes, FosB, c-Fos, Fra-1, c-Jun, JunB, and ATF3 from 1-8 h following OS. The peak level of induced transcription from OS varied from 2- to 128-fold over the first 4 h, depending on the gene and magnitude of OS. Increased transcription at higher levels of OS was also seen for up to 8-h for some of these genes. Protein translation was examined for 24-h following OS using Western blotting methods, and compared to the qPCR responses. We identified six AP-1 family genes that demonstrate quantitative upregulation of expression in response to OS. Two distinct types of quantifiable OS-specific responses were observed; dose-dependent responses, and threshold responses. Our studies show that different levels of OS can regulate the expression of AP-1 transcription factors quantitatively in the human RPE in vitro.",
author = "Edward Chaum and Jinggang Yin and Huaitao Yang and Fridtjof Thomas and Lang, {John C.}",
year = "2009",
month = "12",
day = "15",
doi = "10.1002/jcb.22358",
language = "English (US)",
volume = "108",
pages = "1280--1291",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "6",

}

TY - JOUR

T1 - Quantitative AP-1 gene regulation by oxidative stress in the human retinal pigment epithelium

AU - Chaum, Edward

AU - Yin, Jinggang

AU - Yang, Huaitao

AU - Thomas, Fridtjof

AU - Lang, John C.

PY - 2009/12/15

Y1 - 2009/12/15

N2 - The purpose of this study was to characterize the early molecular responses to quantified levels of oxidative stress (OS) in the human retinal pigment epithelium (RPE). Confluent ARPE-19 cells were cultured for 3 days in defined medium to stabilize gene expression. The cells were exposed to varying levels of OS (0-500 μMH2O2) for 1-8 h and gene expression was followed for up to 24-h after OS. Using real-time qPCR, we quantified the expression of immediate early genes from the AP-1 transcription factor family and other genes involved in regulating the redox status of the cells. Significant and quantitative changes were seen in the expression of six AP-1 transcription factor genes, FosB, c-Fos, Fra-1, c-Jun, JunB, and ATF3 from 1-8 h following OS. The peak level of induced transcription from OS varied from 2- to 128-fold over the first 4 h, depending on the gene and magnitude of OS. Increased transcription at higher levels of OS was also seen for up to 8-h for some of these genes. Protein translation was examined for 24-h following OS using Western blotting methods, and compared to the qPCR responses. We identified six AP-1 family genes that demonstrate quantitative upregulation of expression in response to OS. Two distinct types of quantifiable OS-specific responses were observed; dose-dependent responses, and threshold responses. Our studies show that different levels of OS can regulate the expression of AP-1 transcription factors quantitatively in the human RPE in vitro.

AB - The purpose of this study was to characterize the early molecular responses to quantified levels of oxidative stress (OS) in the human retinal pigment epithelium (RPE). Confluent ARPE-19 cells were cultured for 3 days in defined medium to stabilize gene expression. The cells were exposed to varying levels of OS (0-500 μMH2O2) for 1-8 h and gene expression was followed for up to 24-h after OS. Using real-time qPCR, we quantified the expression of immediate early genes from the AP-1 transcription factor family and other genes involved in regulating the redox status of the cells. Significant and quantitative changes were seen in the expression of six AP-1 transcription factor genes, FosB, c-Fos, Fra-1, c-Jun, JunB, and ATF3 from 1-8 h following OS. The peak level of induced transcription from OS varied from 2- to 128-fold over the first 4 h, depending on the gene and magnitude of OS. Increased transcription at higher levels of OS was also seen for up to 8-h for some of these genes. Protein translation was examined for 24-h following OS using Western blotting methods, and compared to the qPCR responses. We identified six AP-1 family genes that demonstrate quantitative upregulation of expression in response to OS. Two distinct types of quantifiable OS-specific responses were observed; dose-dependent responses, and threshold responses. Our studies show that different levels of OS can regulate the expression of AP-1 transcription factors quantitatively in the human RPE in vitro.

UR - http://www.scopus.com/inward/record.url?scp=71749115655&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=71749115655&partnerID=8YFLogxK

U2 - 10.1002/jcb.22358

DO - 10.1002/jcb.22358

M3 - Article

VL - 108

SP - 1280

EP - 1291

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 6

ER -