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Mitra Alizadeh, Mitsumasa Wada, Claire M. Gelfman, James T. Handa, Leonard M. Hjelmeland; Downregulation of Differentiation Specific Gene Expression by Oxidative Stress in ARPE-19 Cells. Invest. Ophthalmol. Vis. Sci. 2001;42(11):2706-2713.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To investigate how the differentiation of ARPE-19 cells affects the
relative expression of the FGFR genes in
response to oxidative stress.
methods. After differentiation in vitro, APRE-19 cells were treated with t-butyl hydroperoxide (tBH) or hydrogen peroxide
(H2O2) to induce oxidative stress. Viability
and reactive oxygen intermediate (ROI) production were measured using
standard assays. The mRNA expression of FGFR1, FGFR2, cellular retinaldehyde-binding protein
(CRALBP), RPE65, and heme oxygenase-1
(HO-1) were measured by Northern blot analysis as a
function of treatment with tBH and H2O2.
results. ARPE-19 cells were viable at all tBH concentrations tested but showed
progressive loss of viability at concentrations greater than 300 μM
H2O2. Differentiated ARPE-19 cells treated with
tBH or H2O2 resulted in upregulation of the HO-1 and FGFR1 transcripts. The
expression of RPE-differentiated specific genes, including FGFR2, CRALBP, and RPE65 mRNAs, was downregulated with tBH or H2O2 treatment.
conclusions. Oxidative stress in differentiated ARPE-19 cells alters the expression
of FGFR1, FGFR2, CRALBP,
and RPE65 toward levels characteristic of the
undifferentiated state. If similar changes take place in vivo, these
events could alter the proliferative potential, viability, and even the
function of the RPE.
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