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Hironori Matsui, Li-Ren Lin, Dhirendra P. Singh, Toshimichi Shinohara, Venkat N. Reddy; Lens Epithelium-Derived Growth Factor: Increased Survival and Decreased DNA Breakage of Human RPE Cells Induced by Oxidative Stress. Invest. Ophthalmol. Vis. Sci. 2001;42(12):2935-2941.
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purpose. Lens epithelium-derived growth factor (LEDGF) has been shown to be a
growth and survival factor and to be present in a wide variety of cell
types. The purpose of this study was to determine whether LEDGF
enhances survival of human retinal pigment epithelial (RPE) cells when
challenged by oxidative stress or by ultraviolet (UVB) irradiation in a
methods. Primary RPE cells were cultured in standard Dulbecco’s modified
Eagle’s medium (DMEM) containing 15% fetal bovine serum. Protein blot
analysis with antibodies to LEDGF was used to detect LEDGF in RPE
cells. Initially, RPE cells were cultured in the standard medium for 1
day to allow attachment to the culture plates and then cultured in
serum-free DMEM, with and without LEDGF. The trypan blue exclusion
method was used to test RPE cell viability. Single-cell electrophoresis
was used to evaluate single strand breaks of genomic DNA after exposure
to H2O2 or irradiation by UVB.
results. LEDGF was present in RPE cells, predominantly in the nucleus. RPE cells
grew for 1 week and survived for 3 weeks in the presence of LEDGF. In
the absence of LEDGF, they increased in number for the first week and
gradually died in the following 2 weeks. LEDGF protected RPE cells
against H2O2 exposure and UVB irradiation. DNA
damage induced by H2O2 exposure or UVB
irradiation was lower in the presence than in the absence of LEDGF. The
expression of heat shock protein (Hsp)27 was elevated by LEDGF.
conclusions. LEDGF enhanced survival of RPE cells in culture when challenged by
oxidative stress and UVB irradiation. LEDGF protected DNA from
single-strand breakage and upregulated the expression of Hsp27. These
results suggest that LEDGF may be a potential agent for protecting RPE
cells under various stress conditions.
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