Mynampati Bharani Krishna, Sambhav Kumar, Grover Sandeep, Chalam Kakarla V
Department of Ophthalmology, University of Florida, College of Medicine, Jacksonville, FL 32209, USA.
Int J Ophthalmol. 2017 Jan 18;10(1):15-22. doi: 10.18240/ijo.2017.01.03. eCollection 2017.
To evaluate the differential inhibitory effects of bevacizumab on cell proliferation of vascular endothelial growth factor (VEGF)-stimulated choroidal vascular endothelial cells (CVECs) and retinal vascular endothelial cells (RVECs)
VEGF (400 ng/mL) enriched CVECs and RVECs were treated with escalating doses of bevacizumab (0.1, 0.5, 1, 1.5 and 2 mg/mL). Cell proliferation changes were analyzed with WST-1 assay and trypan blue exclusion assay at 48, 72h and 1wk. Morphological changes were recorded with bright field microscopy.
VEGF enriched RVECs showed significantly more decline of cell viability than CVECs after bevacizumab treatment. One week after treatment, RVEC cell proliferation decreased by 29.7%, 37.5%, 52.8%, 35.9% and 45.6% at 0.1, 0.5, 1.0, 1.5 and 2 mg/mL bevacizumab respectively compared to CVEC proliferation decrease of 4.1%, 7.7%, 2.4%, 4.1% and 17.7% (<0.05) by WST-1 assay. Trypan blue exclusion assay also revealed similar decrease in RVEC proliferation of 20%, 60%, 73.3%, 80% and 93.3% compared to CVEC proliferation decrease of 4%, 12%, 22.9%, 16.7% and 22.2% respectively (<0.05). The maximum differential effect between the two cell types was observed at bevacizumab doses of 1.0 and 1.5 mg/mL at all time points. RVECs were 22 fold more sensitive (<0.01) compared to CVECs (52.8% 2.4%) at concentration of 1.0 mg/mL, and 8.7 fold more at 1.5 mg/mL (35.9% 4.1%) 1wk after treatment (<0.05 respectively).
VEGF-enriched RVECs are more susceptible to bevacizumab inhibition than CVECs at clinically used dosage of 1.25 mg and this differential sensitivity between two cell types should be taken into consideration in dosage selection.
评估贝伐单抗对血管内皮生长因子（VEGF）刺激的脉络膜血管内皮细胞（CVECs）和视网膜血管内皮细胞（RVECs）细胞增殖的差异抑制作用
用递增剂量的贝伐单抗（0.1、0.5、1、1.5和2mg/mL）处理富含VEGF（400ng/mL）的CVECs和RVECs。在48、72小时和1周时，用WST-1法和台盼蓝排斥试验分析细胞增殖变化。用明场显微镜记录形态学变化。
贝伐单抗处理后，富含VEGF的RVECs的细胞活力下降明显多于CVECs。处理1周后，通过WST-1试验，与CVEC增殖分别下降4.1%、7.7%、2.4%、4.1%和17.7%相比，RVEC细胞增殖在贝伐单抗浓度为0.1、0.5、1.0、1.5和2mg/mL时分别下降29.7%、37.5%、52.8%、35.9%和45.6%（<0.05）。台盼蓝排斥试验也显示，与CVEC增殖分别下降4%、12%、22.9%、16.7%和22.2%相比，RVEC增殖分别下降20%、60%、73.3%、80%和93.3%（<0.05）。在所有时间点，两种细胞类型之间的最大差异效应在贝伐单抗剂量为1.0和1.5mg/mL时观察到。处理1周后，在浓度为1.0mg/mL时，RVECs的敏感性比CVECs高22倍（<0.01）（52.8%对2.4%），在1.5mg/mL时高8.7倍（35.9%对4.1%）（均<0.05）。
在临床使用剂量1.25mg时，富含VEGF的RVECs比CVECs对贝伐单抗抑制更敏感，在剂量选择时应考虑两种细胞类型之间的这种差异敏感性。
