阿柏西普对葡萄膜黑色素瘤细胞的作用及其机制研究  

Effect and mechanism of aflibercept on uveal melanoma cells

作　　者：孙浩 李姣[3] 毕宏生[3] 王兴荣[3] SUN Hao;LI Jiao;BI Hongsheng;WANG Xingrong(Shandong University of Traditional Chinese Medicine,Jinan 250002,Shandong Province,China;Lanling County People’s Hospital of Linyi City,Linyi 276000,Shandong Province,China;Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine,Shandong Institute of Eye Disease Control,Jinan 250002,Shandong Province,China)

机构地区：[1]山东中医药大学,山东省济南市250002 [2]兰陵县人民医院,山东省临沂市276000 [3]山东中医药大学附属眼科医院、山东省眼病防治研究院,山东省济南市250002

出　　处：《眼科新进展》2022年第4期284-288,共5页Recent Advances in Ophthalmology

基　　金：山东省医药卫生科技发展计划项目(编号2018ws194);山东省自然科学基金面上项目(编号ZR2020MH393)。

摘　　要：目的探索阿柏西普对葡萄膜黑色素瘤细胞的作用及其机制。方法将B16F10细胞(葡萄膜黑色素瘤细胞)分为实验组和对照组,对照组不使用药物处理,实验组分别用0.2 g·L^(-1)、2.0 g·L^(-1)、4.0 g·L^(-1)阿柏西普进行干预。通过实时细胞电子分析系统(RT-CES)、CCK-8法探讨不同剂量的阿柏西普对葡萄膜黑色素瘤细胞生长过程的影响;ELISA分析阿柏西普对葡萄膜黑色素瘤细胞中VEGF-A含量的影响;RT-PCR检测不同剂量阿柏西普对葡萄膜黑色素瘤细胞VEGF-A mRNA表达的影响;流式细胞仪分析不同剂量阿柏西普对葡萄膜黑色素瘤细胞周期和凋亡的影响。结果0.2 g·L^(-1)、2.0 g·L^(-1)、4.0 g·L^(-1)的阿柏西普作用B16F10细胞24 h后,细胞存活率分别下降至(95.10±1.76)%、(87.33±2.20)%和(74.36±1.67)%。RT-PCR检测结果显示阿柏西普能够下调细胞中VEGF-A mRNA的表达;ELISA检测结果表明,阿柏西普可抑制细胞中VEGF-A的水平,其中4.0 g·L^(-1)阿柏西普抑制作用最为明显。此外,与对照组[细胞凋亡率0%,G1期细胞比例为(27.60±0.36)%]相比,0.2 g·L^(-1)、2.0 g·L^(-1)、4.0 g·L^(-1)阿柏西普干预24 h后,B16F10细胞凋亡率分别升高为3.51%、11.10%和14.77%,G1期细胞比例分别升高至(64.09±0.34)%、(66.02±0.65)%、(67.49±0.33)%,差异均有统计学意义(均为P<0.05)。结论阿柏西普对葡萄膜黑色素瘤细胞有显著抑制作用,其作用机制为抑制细胞中VEGF-A水平,从而诱导细胞发生凋亡和S期阻滞,其作用具有浓度依赖性。Objective To investigate the effect and mechanism of aflibercept on uveal melanoma cells.Methods B16F10 cells(uveal melanoma cells)were divided into the experimental group and control group.Cells in the control group were not treated with drugs,while cells in the experimental group were intervened with 0.2 g·L^(-1),2.0 g·L^(-1),and 4.0 g·L^(-1) aflibercept.Real-time cell electronic sensing(RT-CES)and CCK-8 assays were used to explore the effect of different doses of aflibercept on the growth process of uveal melanoma cells.ELISA assay was performed to investigate the effect of aflibercept on the content of vascular endothelial growth factor A(VEGF-A)in uveal melanoma cells,real-time polymerase chain reaction(RT-PCR)assay was performed to investigate the effects of different doses of aflibercept on the mRNA expression of VEGF-A in uveal melanoma cells,and flow cytometry was performed to investigate the effects of different doses of aflibercept on cycle and apoptosis of uveal melanoma cells.Results It was found that the survival rates of B16F10 cells treated by 0.2 g·L^(-1),2.0 g·L^(-1) and 4.0 g·L^(-1) of aflibercept for 24 h fell to(95.10±1.76)%,(87.33±2.20)%and(74.36±1.67)%,respectively.RT-PCR assay results showed that aflibercept down-regulated the mRNA expression of VEGF-A in cells.ELISA assay results showed that aflibercept inhibited VEGF-A levels in cells,with 4.0 g·L^(-1) aflibercept having the most significant inhibitory effect.In addition,compared with the control group apoptosis rate 0%,proportion of cells in G1 phase(27.60±0.36)%,the apoptosis rates of B16F10 cells in the experimental group increased to 3.51%,11.10%and 14.77%after being intervened by 0.2 g·L^(-1),2.0 g·L^(-1) and 4.0 g·L^(-1) aflibercept for 24 h,respectively,and the proportion of cells in G1 phase increased to(64.09±0.34)%,(66.02±0.65)%and(67.49±0.33)%,with statistically significant differences(all P<0.05).Conclusion Aflibercept shows significant inhibitory effects on uveal melanoma cells by inhibiting the VEGF-A level

关 键 词：阿柏西普 葡萄膜黑色素瘤 VEGF 作用机制 

分 类 号：R773.9[医药卫生—眼科]