盐酸埃克替尼诱导NF2神经鞘瘤细胞增殖和凋亡研究  

Study on icotinib of the proliferation and apoptosis of neurofibromatosis type 2 related schwannoma cells

作　　者：汪颖[1] 赵赋[1] 张晶[1] 王博[2] 张琪[1] 孙异临[1] 刘丕楠[1,2] 

机构地区：[1]首都医科大学北京神经外科研究所,北京100050 [2]首都医科大学附属北京天坛医院,北京100050

出　　处：《东北农业大学学报》2015年第7期57-61,共5页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(81372715)

摘　　要：探讨盐酸埃克替尼对2型神经纤维瘤病（NF2）神经鞘瘤细胞增殖和凋亡的影响。运用0～80μmol·L^-1盐酸埃克替尼处理NF2听神经鞘瘤原代细胞及大鼠神经鞘瘤细胞系RT4,孵育48 h后通过CCK8比色法检测药物对细胞增殖活力的影响;流式细胞仪检测药物对细胞凋亡的影响;电子显微镜观察药物处理后细胞显微结构变化;同时应用免疫印迹法及实时定量PCR观察凋亡相关蛋白表达情况。盐酸埃克替尼作用后,细胞增殖活性显著降低,呈剂量依赖性。流式细胞仪检测,10、20、40、80μmol·L^-1盐酸埃克替尼作用48 h后,凋亡率分别为（6.10±0.35）%、（10.52±0.87）%、（11.78±0.63）%、（16.05±1.02）%。免疫印迹检测发现,半胱氨酸蛋白水解酶3（Caspase-3）剪切体增多,实时定量PCR结果显示bcl-2基因表达降低,bax基因表达增加。结果表明,盐酸埃克替尼能通过活化Caspase-3及调控bcl-2、bax基因表达诱导细胞凋亡而抑制神经鞘瘤细胞增殖。To explore the role of icoUnib on the proliferation and apoptosis of neurofibromatosis type 2 （NF2） related schwannoma cells. Primary cultured NF2 related human schwannoma cells and rat schwannoma cell line RT4 cells were treated in vitro with icotinib at a concentration gradient of 0-80 μmol· L^-1, and its role on the cell proliferation and apoptosis were analyzed by the CCK8 assay and flow cytometer, respectively. The ultrastructural changes was evaluated by electron microscopy, and apoptosis-related proteins and genes were detected using Western-blot and RT-PCR. Results showed that icotinib inhibites the proliferation of primary NF2-related schwannoma cells and RT4 cells in a dose dependent manner, after treatment with 10, 20, 40 and 80μmol· L^-1 icotinib for 48 h, the rates of cell apoptosis were （6.10±0.35）%,（10.52 ± 0.87）%, （11.78 ± 0.63）% and （16.05± 1.02）%, respectively. These effects were associated with an increased expression of cleaved-Caspase-3 and altered expression of bcl-2, bax genes. Thus, icotinib may plays a role in inhibiting proliferation, and increasing apoptosis of schwannoma cells.

关 键 词：盐酸埃克替尼 2型神经纤维瘤病 神经鞘瘤 表皮生长因子受体 

分 类 号：R739.4[医药卫生—肿瘤]