Effects of Bevacizumab on Bcl-2 Expression and Apoptosis in Retinal Pigment Epithelial Cells under Oxidative Stress
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Purpose: To evaluate the effects of bevacizumab on expression of B-cell leukemia/lymphoma (Bcl)-2 and apoptosisin retinal pigment epithelial (RPE) cells under oxidative stress conditions. Methods: RPE cells were treated with H2O2 (0, 100, 200, 300, and 400 μM) and bevacizumab at or above thedoses normally used in clinical practice (0, 0.33, 0.67, 1.33, and 2.67 mg/mL). Cell apoptosis was measuredusing flow cytometry with annexin V-fluorescein isothiocyanate. The expression of Bcl-2 mRNA was determinedusing reverse transcription polymerase chain reaction. Results: Under low oxidative stress conditions (H2O2 100 μM), cell apoptosis was not significantly different atany concentration of bevacizumab, but Bcl-2 mRNA expression decreased with increasing concentration ofbevacizumab (0.33, 0.67, 1.33, and 2.67 mg/mL). Under moderate oxidative stress conditions (H2O2 200 μM),Bcl-2 mRNA expression decreased with increasing concentration of bevacizumab (0.33, 0.67, 1.33, and 2.67mg/mL), but cell apoptosis increased only at 2.67 mg/mL of bevacizumab. Under high oxidative stress (300μM) conditions, cell apoptosis increased at high concentrations of bevacizumab (1.33 and 2.67 mg/mL), but itdid not correlate with Bcl-2 expression. Conclusions: Withdrawal of vascular endothelial growth factor can lead to RPE cell apoptosis and influencesthe expression of anti-apoptotic genes such as Bcl-2 under oxidative stress conditions. Since oxidative stresslevels of each patient are unknown, repeated injections of intravitreal bevacizumab, as in eyes with age-relatedmacular degeneration, might influence RPE cell survival.
Abstract
Purpose: To evaluate the effects of bevacizumab on expression of B-cell leukemia/lymphoma (Bcl)-2 and apoptosisin retinal pigment epithelial (RPE) cells under oxidative stress conditions. Methods: RPE cells were treated with H2O2 (0, 100, 200, 300, and 400 μM) and bevacizumab at or above thedoses normally used in clinical practice (0, 0.33, 0.67, 1.33, and 2.67 mg/mL). Cell apoptosis was measuredusing flow cytometry with annexin V-fluorescein isothiocyanate. The expression of Bcl-2 mRNA was determinedusing reverse transcription polymerase chain reaction. Results: Under low oxidative stress conditions (H2O2 100 μM), cell apoptosis was not significantly different atany concentration of bevacizumab, but Bcl-2 mRNA expression decreased with increasing concentration ofbevacizumab (0.33, 0.67, 1.33, and 2.67 mg/mL). Under moderate oxidative stress conditions (H2O2 200 μM),Bcl-2 mRNA expression decreased with increasing concentration of bevacizumab (0.33, 0.67, 1.33, and 2.67mg/mL), but cell apoptosis increased only at 2.67 mg/mL of bevacizumab. Under high oxidative stress (300μM) conditions, cell apoptosis increased at high concentrations of bevacizumab (1.33 and 2.67 mg/mL), but itdid not correlate with Bcl-2 expression. Conclusions: Withdrawal of vascular endothelial growth factor can lead to RPE cell apoptosis and influencesthe expression of anti-apoptotic genes such as Bcl-2 under oxidative stress conditions. Since oxidative stresslevels of each patient are unknown, repeated injections of intravitreal bevacizumab, as in eyes with age-relatedmacular degeneration, might influence RPE cell survival.
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