Effect of Anti-vascular Endothelial Growth Factor Antibody on the Survival of Cultured Retinal Ganglion Cells

Korean J Ophthalmol. 2017 Aug;31(4):360-365. doi: 10.3341/kjo.2017.0054. Epub 2017 Jul 11.

Abstract

Purpose: To investigate the effects of anti-vascular endothelial growth factor (VEGF) antibody on the survival of retinal ganglion cell (RGC)-5 cells differentiated with staurosporine under oxidative stress.

Methods: We used real-time polymerase chain reaction and Western blot to confirm the expression of VEGF, VEGF receptor (VEGFR)-1 and VEGFR-2 in RGC-5 cells differentiated with staurosporine for 6 hours. The differentiated RGC-5 cells were treated with 800 μM hydrogen peroxide (H₂O₂) for 24 hours to induce oxidative stress. Then, the survival rate of RGC-5 was confirmed by lactate dehydrogenase assay at each concentration (0, 0.01, 0.1, and 1 mg) using bevacizumab as the anti-VEGF antibody. The expression of VEGF, VEGFR-1, and VEGFR-2 was confirmed using real-time polymerase chain reaction.

Results: VEGF, VEGFR-1, and VEGFR-2 were all expressed in differentiated RGC-5 cells. When RGC-5 cells were simultaneously treated with bevacizumab and 800 μM H₂O₂, survival of RGC-5 decreased with bevacizumab concentration. VEGF expression in RGC-5 cells increased with increasing concentration of bevacizumab. Similar patterns were observed for VEGFR-1 and VEGFR-2, but the degree of increase was smaller than that for VEGF.

Conclusions: When bevacizumab was administered to differentiated RGC-5 cells, the cell damage caused by oxidative stress increased. Therefore, given these in vitro study results, caution should be exercised with bevacizumab treatment.

Keywords: Anti-vascular endothelial growth factor; Bevacizumab; Oxidative stress; RGC-5; Retinal ganglion cell.

MeSH terms

  • Angiogenesis Inhibitors / pharmacology
  • Animals
  • Antibodies / pharmacology*
  • Bevacizumab / pharmacology
  • Blotting, Western
  • Cell Differentiation
  • Cell Survival
  • Cells, Cultured
  • Gene Expression Regulation / drug effects
  • Mice
  • Oxidative Stress / genetics*
  • RNA / genetics*
  • Real-Time Polymerase Chain Reaction
  • Retinal Ganglion Cells / cytology*
  • Vascular Endothelial Growth Factor A / biosynthesis
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / immunology*

Substances

  • Angiogenesis Inhibitors
  • Antibodies
  • Vascular Endothelial Growth Factor A
  • Bevacizumab
  • RNA