TY - JOUR
T1 - Effects of bevacizumab on endoplasmic reticulum stress in hypoxic retinal pigment epithelial cells
AU - Park, Joo Hee
AU - Kim, Moosang
AU - Oh, Jong Hyun
N1 - Publisher Copyright:
© 2017 Park et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2017/6
Y1 - 2017/6
N2 - Purpose: To investigate the effects of bevacizumab on endoplasmic reticulum (ER) stress in human retinal pigment epithelial (RPE) cells cultured under hypoxic conditions. Methods: RPE cells (ARPE-19) were cultured under hypoxic conditions (1% O2) with or without bevacizumab (0.3125 mg/mL) for 24 and 48 h. Cell viability was measured by a PrestoBlue assay. The expression of vascular endothelial growth factor (VEGF), binding protein/glucose-regulated protein 78 (BiP/GRP78), and C/EBP homologous protein-10 (CHOP) mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). BiP/GRP78 and CHOP protein levels in the cells were assessed by western blot. VEGF protein in the media was quantified by enzyme-linked immunosorbent assay (ELISA). Results: Under hypoxic conditions, cell viability decreased and mRNA and protein levels of VEGF, BiP/GRP78, and CHOP increased compared to those under normoxic conditions. Bevacizumab improved cell viability and reduced the expression of VEGF mRNA under hypoxic conditions. Bevacizumab also reduced the expression of both mRNA and protein of two ER stress indicators, BiP/GRP78 and CHOP, under hypoxic conditions. Conclusions: Bevacizumab mitigated ER stress in human RPE cells cultured under hypoxic conditions. This effect may be involved in the improved cell viability and reduction of VEGF expression after bevacizumab treatment of hypoxic RPE cells in vitro. However, the effects of bevacizumab on RPE cells under experimental conditions are unlikely to be clinically equivalent to those in the human eye.
AB - Purpose: To investigate the effects of bevacizumab on endoplasmic reticulum (ER) stress in human retinal pigment epithelial (RPE) cells cultured under hypoxic conditions. Methods: RPE cells (ARPE-19) were cultured under hypoxic conditions (1% O2) with or without bevacizumab (0.3125 mg/mL) for 24 and 48 h. Cell viability was measured by a PrestoBlue assay. The expression of vascular endothelial growth factor (VEGF), binding protein/glucose-regulated protein 78 (BiP/GRP78), and C/EBP homologous protein-10 (CHOP) mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). BiP/GRP78 and CHOP protein levels in the cells were assessed by western blot. VEGF protein in the media was quantified by enzyme-linked immunosorbent assay (ELISA). Results: Under hypoxic conditions, cell viability decreased and mRNA and protein levels of VEGF, BiP/GRP78, and CHOP increased compared to those under normoxic conditions. Bevacizumab improved cell viability and reduced the expression of VEGF mRNA under hypoxic conditions. Bevacizumab also reduced the expression of both mRNA and protein of two ER stress indicators, BiP/GRP78 and CHOP, under hypoxic conditions. Conclusions: Bevacizumab mitigated ER stress in human RPE cells cultured under hypoxic conditions. This effect may be involved in the improved cell viability and reduction of VEGF expression after bevacizumab treatment of hypoxic RPE cells in vitro. However, the effects of bevacizumab on RPE cells under experimental conditions are unlikely to be clinically equivalent to those in the human eye.
UR - http://www.scopus.com/inward/record.url?scp=85020406376&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0179048
DO - 10.1371/journal.pone.0179048
M3 - Article
C2 - 28591217
AN - SCOPUS:85020406376
SN - 1932-6203
VL - 12
JO - PLoS ONE
JF - PLoS ONE
IS - 6
M1 - e0179048
ER -