Jun Jong Hwa, Sohn Wern-Joo, Lee Youngkyun, Kim Jae-Young
Department of Ophthalmology, School of Medicine, Dongsan Medical Center, Keimyung University.
Department of Oral Biochemistry, School of Dentistry, IHBR, Kyungpook National University, Daegu, South Korea.
Clin Ophthalmol. 2016 Jun 27;10:1167-74. doi: 10.2147/OPTH.S103443. eCollection 2016.
The molecular and cellular effects of anti-vascular endothelial growth factor monoclonal antibody (bevacizumab) on lens epithelial cells (LECs) were examined using both an immortalized human lens epithelial cell line and a porcine capsular bag model. After treatment with various concentrations of bevacizumab, cell viability and proliferation patterns were evaluated using the water-soluble tetrazolium salt assay and 5-bromo-2'-deoxyuridine enzyme-linked immunosorbent assay, respectively. The scratch assay and Western blot analysis were employed to validate the cell migration pattern and altered expression levels of signaling molecules related to the epithelial-mesenchymal transition (EMT). Application of bevacizumab induced a range of altered cellular events in a concentration-dependent manner. A 0.1-2 mg/mL concentration demonstrated dose-dependent increase in proliferation and viability of LECs. However, 4 mg/mL decreased cell proliferation and viability. Cell migrations displayed dose-dependent retardation from 0.1 mg/mL bevacizumab treatment. Transforming growth factor-β2 expression was markedly increased in a dose-dependent manner, and α-smooth muscle actin, matrix metalloproteinase-9, and vimentin expression levels showed dose-dependent changes in a B3 cell line. Microscopic observation of porcine capsular bag revealed changes in cellular morphology and a decline in cell density compared to the control after 2 mg/mL treatment. The central aspect of posterior capsule showed delayed confluence, and the factors related to EMT revealed similar expression patterns to those identified in the cell line. Based on these results, bevacizumab modulates the proliferation and viability of LECs and induces morphological alterations through the modulation of expression patterns of specific factors related to the EMT.
使用永生化人晶状体上皮细胞系和猪囊袋模型，研究了抗血管内皮生长因子单克隆抗体（贝伐单抗）对晶状体上皮细胞（LECs）的分子和细胞效应。用不同浓度的贝伐单抗处理后，分别使用水溶性四氮唑盐法和5-溴-2'-脱氧尿苷酶联免疫吸附测定法评估细胞活力和增殖模式。采用划痕试验和蛋白质印迹分析来验证细胞迁移模式以及与上皮-间质转化（EMT）相关的信号分子表达水平的变化。贝伐单抗的应用以浓度依赖性方式诱导了一系列细胞事件的改变。0.1-2mg/mL的浓度显示LECs的增殖和活力呈剂量依赖性增加。然而，4mg/mL降低了细胞增殖和活力。从0.1mg/mL贝伐单抗处理开始，细胞迁移显示出剂量依赖性延迟。在B3细胞系中，转化生长因子-β2的表达以剂量依赖性方式显著增加，α-平滑肌肌动蛋白、基质金属蛋白酶-9和波形蛋白的表达水平显示出剂量依赖性变化。猪囊袋的显微镜观察显示，与2mg/mL处理后的对照相比，细胞形态发生了变化，细胞密度下降。后囊中央部分显示融合延迟，与EMT相关的因子显示出与细胞系中鉴定的相似表达模式。基于这些结果，贝伐单抗通过调节与EMT相关的特定因子的表达模式来调节LECs的增殖和活力，并诱导形态学改变。
