雷珠单抗选择性体外抑制睫状体恶性黑色素瘤  

Ranibizumab selective inhibiting proliferation of malignant melanoma of ciliary body in vitro

作　　者：李姣[1] 王兴荣[1] 崔彦[2] 刘正峰[3] 王影[3] 李洋[1] 

机构地区：[1]山东中医药大学附属眼科医院,山东省济南市250002 [2]山东中医药大学附属眼科医院眼科研究所山东省高校中西医结合眼病防治技术重点实验室,山东省济南市250002 [3]山东中医药大学第二临床学院,山东省济南市250014

出　　处：《眼科新进展》2013年第11期1021-1025,共5页Recent Advances in Ophthalmology

基　　金：山东省自然科学基金(编号:Y2006C101)~~

摘　　要：目的探讨雷珠单抗是否能抑制恶性黑色素瘤细胞的生长并评估其在体外对人视网膜色素上皮细胞的安全性。方法人视网膜色素上皮细胞系(human retinal pigmental epithelial cell line,ARPE-19)和临床获得的恶性黑色素瘤细胞均分为空白对照组、血管内皮生长因子(vascular endothelial growth factor,VEGF)处理组和雷珠单抗处理组。VEGF处理组预设VEGF浓度分别为1μg·L-1、5μg·L-1、10μg·L-1和20μg·L-1,雷珠单抗处理组预设雷珠单抗的浓度分别为0.10 g·L-1、0.25 g·L-1、0.50 g·L-1和1.00 g·L-1。采用实时细胞电子分析系统(RT-CES)分别监测各组细胞的动态变化,ELISA法检测对照组和雷珠单抗组VEGF的表达水平。结果与空白对照(0μg·L-1VEGF)组相比,10μg·L-1和20μg·L-1VEGF可促进恶性黑色素瘤细胞的生长(均为P<0.05)。但是同样浓度的VEGF作用下ARPE-19的生存率与对照组相比没有明显增长(均为P>0.05)。当加入0.25 g·L-1雷珠单抗时,恶性黑色素瘤细胞的生存率下降至(42.5±5.0)%(P<0.01)。0.50 g·L-1和1.00 g·L-1雷珠单抗作用于ARPE-19时,细胞生存率分别是(55.6±1.1)%和(42.8±4.0)%。与空白对照(0 g·L-1雷珠单抗)组恶性黑色素瘤细胞表达的VEGF水平(1533.4±7.0)ng·L-1相比,0.10 g·L-1、0.25 g·L-1、0.50 g·L-1和1.00 g·L-1雷珠单抗处理后其VEGF水平分别下降到(822.6±9.3)ng·L-1、(875.0±7.8)ng·L-1、(836.1±6.3)ng·L-1和(844.8±6.0)ng·L-1(均为P<0.05)。正常的ARPE-19可表达(908.8±9.1)ng·L-1VEGF,而经不同浓度雷珠单抗处理后,其表达的VEGF水平与对照组相比差异均无统计学意义(均为P>0.05)。结论雷珠单抗选择性抑制恶性黑色素瘤细胞的增殖,而对ARPE-19的作用较弱。Objective To delineate whether ranibizumab can inhibit malignant melanoma growth and evaluate safety of ranibizumab on human retinal pigment epitheli- al cells in vitro. Methods Human retinal pigment epithelial cell line（ARPE-19） and malignant melanoma cells obtained from clinic were divided into three groups： control group ,vascular endothelial growth factor（VEGF） group and ranibizumab group ,respectively. Concentrations of VEGF with 1μg·L^-1 ,5μg·L^-1,10 μg·L^-1 and 20 μg·L^-1 were given in VEGF group, and concentrations of ranibizumab with 0. 10 g ~ L-1, 0. 25 g ~ L- 1,0.50 g ~ L- 1 and 1.00 g ~ L- 1 in ranibizumab group. The real-time cell electronic sensing（RT-CES） assay was utilized to monitor the dynamic changes of cell in three groups in vitro. ELISA method was used to assay the expression of VEGF in control group and ranibizumab group. Results Compared with control group （0μg·L^-1 VEGF） ,10μg·L^-1 and 20μg·L^-1 VEGF could inhibit the growth of malignant mela- noma cells （ all P 〈 0. 05 ）. However, there was no statistical difference in growth of ARPE-19 under same concentration of VEGF between two groups（ all P 〉 0.05 ）. When ranibizumab was 0.25 g·L^-1, the survival rate of malignant melanoma cells decreased to（42.5 ±5.0）% （P〈0.01）. When ranibizumab was 0.50g·L^-1 and 1.00g·L^-1 ,the survival rate of ARPE-19 were（55.6 ±1. 1）% and（42.8 ±4.0）% ,respectively. When ranibizumab were 0. 10g·L^-1 ,0.25 g·L^-1 ,0.50g·L^-1 and 1.00g·L^-1,the levels of VEGF of malignant melanoma cells were declined to （ 822.5 ± 9.3 ）ng·L^-1, （ 875.0 ± 7. 8）ng·L^-1 , （836.1±6.3）ng·L^-1 and（844.8 ±6.0） ng·L^-1 ,respectively. There were significant differences between the above groups and control group （ 1533.4 ± 7.0 ）g·L^-1 （ all P 〈 0.05 ）. In addition, normal ARPE-19 could express（908.75 ± 9.10） ng·L^-1 VEGF,while there was no significant differences between ranibizumab groups and control group（ all P 〉 0.05 ）.

关 键 词：雷珠单抗 血管内皮生长因子 恶性黑色素瘤细胞 人视网膜色素上皮细胞 实时细胞电子分析系统 

分 类 号：R730.43[医药卫生—肿瘤]