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Jong Hwa Jun, Wern-Joo Sohn, Youngkyun Lee, Sung Dong Chang, Jae-Young Kim; The Non-antiangiogenic Effects of Anti-Vascular Endothelial Growth Factor Monoclonal Antibody (Bevacizumab) on Lens Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):734.
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The vascular endothelial growth factor (VEGF) was identified in the lens fiber and lens epithelial cells through the embryonic development to postnatal period. This study evaluated the cellular effects of anti-VEGF monoclonal antibody (bevacizumab) on lens epithelial cells (LEC). In this study, we evaluated non-antiangiogenic effecs of bevacizumab on both cultured immortalized human lens epithelial cells (HLE-B3) and in vitro porcine lens capsular bag.
To analysis the cell viability and proliferation after treatment of various concentrations of bevacizumab, cell counting kit-8 (CCK-8) assay and 5-brome-2’-deoxyuridine enzyme linked immunosorbent assay were employed respectively. In addition, scratch assay and immunologic evaluation were also employed to validate the cell migration and the alterations of expression pattern in specific factors, such as transforming growth factor (TGF)-β2 and α-smooth muscle actin (SMA) after treatment of various concentrations of bevacizumab.
Overall, the applications of bevacizumab induced a range of altered cellular events with concentration specific manner. Bevacizumab treatment showed cellular cytotoxic effect and decreased proliferations after 4 mg/mL concentration. Especially, 0.1 mg/mL concentration of bevacizumab treatment showed the time-dependent toxicity. Apparent decrement of cell migration was identified over 2 mg/mL concentration. TGF-β2 expression was markedly increased and α-SMA was decreased with dose-dependent manner in immunoblot analysis. Immunostaining of α-SMA also showed the decreased localization patterns and microscopic observation revealed change of LEC morphology with decline of cell density in porcine lens capsular bag after 2 mg/mL treatment.
Based on these results, bevacizumab treatment induced cytotoxic effects and impeded cell migration and proliferation in HLE-B3 cells. Furthermore, bevacizumab induces morphological alterations and modulation of expression in specific factors that were related to the epithelial mesenchymal transition.
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