Effects of Bevacizumab, Ranibizumab, and Aflibercept on MicroRNA Expression in a Retinal Pigment Epithelium Cell Culture Model of Oxidative Stress.

PURPOSE

This study aimed to evaluate the effects of bevacizumab, ranibizumab, and aflibercept on the microRNA (miRNA) expression in human retinal pigment epithelium cell (ARPE-19) culture model of oxidative stress.

METHODS

Control cells were cultured in the hydrogen peroxide (HO)-free medium. In HO group ARPE-19 cells were exposed to 600 μM HO alone for 18 h. In study groups, cells were preincubated with bevacizumab, ranibizumab, and aflibercept (1.25-2.5, 0.5 and 2.0 mg/mL, respectively) for 3 h before HO exposure. Another group of ARPE-19 cells were incubated with drugs for 3 h without HO exposure. Cell viability and vascular endothelial growth factor (VEGF) levels were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and enzyme-linked immunosorbent assay. The expression levels of 1,152 miRNAs were determined by quantitative real-time PCR.

RESULTS

Incubation with 600 μM HO alone for 18 h decreased cell viability by ∼50%. Cell viability was greater in the anti-VEGF drug groups compared with the HO group, but the differences were not significant (P > 0.05). VEGF levels were significantly lower in the anti-VEGF drug groups compared with the HO group (P < 0.05 for all study groups), with no significant differences between the study groups (P > 0.05). Incubation with anti-VEGF drugs alone had no effect on miRNA expression in ARPE-19 cells. However, preincubation with bevacizumab, ranibizumab, and aflibercept significantly altered the profile of HO-modulated miRNA expression.

CONCLUSIONS

Preincubation with anti-VEGF drugs can alter the miRNA expression profile in response to HO-induced oxidative stress, and these drugs may have epigenetic effects.