Sharma Rajesh K, Chalam Kakarla V
Department of Ophthalmology, University of Florida, College of Medicine, Jacksonville, Florida 32209, USA.
Acta Ophthalmol. 2009 Sep;87(6):618-22. doi: 10.1111/j.1755-3768.2008.01410.x. Epub 2009 Jun 26.
The effects of bevacizumab on cell viability and proliferation in a commonly used retinal ganglion cell line, RGC-5, were examined.
RGC-5 cells were exposed to 0.1 mg/ml, 1 mg/ml and 2 mg/ml of commercially available bevacizumab in vitro. To examine the specificity of effects, cells were also cultured with increasing and comparable concentrations of proteins (increasing the concentration of proteins in the culture media by 0.1 mg/ml, 1 mg/ml and 2 mg/ml by using additional fetal bovine serum [FBS] and bovine serum albumin [BSA]). Cell proliferation was assessed using a WST-1 kit, crystal violet staining and bromodeoxyuridine (BrdU) incorporation. Cytotoxic effects were assessed by quantifying cell numbers in proliferation-deficient RGC-5 following exposure to bevacizumab using the WST-1 kit, microscopic examination of cells stained with propidium iodide (PI) cells and flow cytometry for differential staining with PI.
Bevacizumab was not toxic to RGC-5 cells in the tested concentrations. It had a stimulatory effect on cell proliferation. A stimulatory effect on proliferation was also noted when equivalent amounts of proteins from FBS or BSA were used, which suggests that bevacizumab may stimulate proliferation non-specifically by increasing the protein contents of the cell growth environment.
Results suggest that intravitreal injection of bevacizumab could alter the internal milieu of the eye by increasing protein concentrations to elicit functional responses in retinotypic cells. This may be especially relevant for cells outwith the control of vascular endothelial growth factor.
研究贝伐单抗对常用视网膜神经节细胞系RGC - 5细胞活力和增殖的影响。
将RGC - 5细胞在体外分别暴露于0.1 mg/ml、1 mg/ml和2 mg/ml的市售贝伐单抗。为了检验作用的特异性,还将细胞与浓度递增且相当的蛋白质一起培养(通过添加额外的胎牛血清[FBS]和牛血清白蛋白[BSA],使培养基中蛋白质浓度分别增加0.1 mg/ml、1 mg/ml和2 mg/ml)。使用WST - 1试剂盒、结晶紫染色和溴脱氧尿苷(BrdU)掺入法评估细胞增殖。通过使用WST - 1试剂盒在贝伐单抗处理后对增殖缺陷的RGC - 5细胞进行细胞计数、用碘化丙啶(PI)染色的细胞显微镜检查以及PI差异染色的流式细胞术来评估细胞毒性作用。
在测试浓度下,贝伐单抗对RGC - 5细胞无毒。它对细胞增殖有刺激作用。当使用等量来自FBS或BSA的蛋白质时,也观察到了对增殖的刺激作用,这表明贝伐单抗可能通过增加细胞生长环境中的蛋白质含量非特异性地刺激增殖。
结果表明,玻璃体内注射贝伐单抗可能通过增加蛋白质浓度来改变眼内环境,从而在视网膜型细胞中引发功能反应。这对于不受血管内皮生长因子控制的细胞可能尤其相关。