去甲斑蝥素通过线粒体途径诱导胶质瘤细胞凋亡的研究  

Norcantharidin induces apoptosis in human glioma cell via mitochondrial pathway

作　　者：庞长河[1] 焦红亮[1] 杨凤东[1] 刘宇[1] 宋振宇[1] 杜伟[1] Pang Changhe;Jiao Hongliang;Yang Fengdong;Liu Yu;Song Zhenyu;Du Wei(Department of Neurosurgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Chin)

机构地区：[1]郑州大学第一附属医院神经外科,450052

出　　处：《中华实验外科杂志》2018年第5期812-814,共3页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关项目（201602083）

摘　　要：目的 探讨去甲斑蝥素（NCTD）诱导胶质瘤细胞凋亡的分子机制.方法 不同浓度NCTD（25、50、100、200 μmol/L）分别处理胶质瘤C6细胞24、48 h,噻唑蓝（MTT）法检测药物对细胞的生长抑制作用;膜联蛋白V-异硫氰酸荧光素（Annexin V-FITC）/碘化丙锭（PI）染色法检测NCTD对C6细胞凋亡的影响;Western blot法检测B细胞淋巴瘤/白血病-2（bcl-2）家族蛋白的表达.结果 MTT法检测显示NCTD呈时间、剂量性抑制细胞增殖,处理24、48 h后半数抑制浓度（IC50）分别为96.5、48.7 μmol/L,对细胞增殖具有明显抑制作用（F=78.202、897.714,P=0.000）;流式细胞仪结果显示,10、30、50 μmol/L NCTD作用C6细胞10 h,细胞凋亡率分别（2.0±2.9）％、（8.4±4.1）％、（19.7±5.7）％,与对照组[（0.9±3.1）％]比较,差异有统计学意义（F=76.246,P=0.000）;Western blot结果显示,25、50、100 μmol/L处理C6细胞24h后,细胞Bcl-2、Mcl-1蛋白的表达下调.结论 NCTD具有较强的抑制胶质瘤细胞生存和诱导凋亡作用,作用机制与通过诱导bcl-2家族抗凋亡蛋白表达下调,激活内源性线粒体凋亡通路有关.Objective To elucidate the molecular mechanism of norcantharidin （NCTD） in inducing apoptosis of glioma cells.Methods Glioma C6 cells were treated with various dosages of NCTD （25,50,100,200 μmol/L） for 24,48 h.The cell growth inhibition was measured by methyl thiazol tetrazolium（MTT） assay.The apoptosis of cells were analyzed by Annexin V-fluoresceine isothiocyanate （FITC）/propidium iodide （PI） staining.The signaling pathway changes were detected by Western blotting.Results NCTD effectively inhibited cell proliferation of C6 cells in a time and dose-dependent manner.The 24 h and 48 h half maximal inhibitory concentration （IC50） were 96.5 and 48.7 μmol/L.It had a significant inhibition of cell proliferation at 24 h and 48 h （F=78.202,897.714,P =0.000）.Compared with the control group （0.9 ±3.1）%,after 10 h treatment,the cell apoptosis rates induced by 10,30,50 μmol/L NCTD were （2.0 ± 2.9） %,（8.4 ± 4.1） % and （19.7 ± 5.7） %,respectively,which indicated that NCTD could induce apoptosis of cells in a dose-dependent manner （F =76.246,P =0.000）.Meanwhile,the expression levels anti-apoptotic proteins B cell lymphoma/leukemia-2 （bcl-2） and Mcl-1 were significantly reduced.Conclusion NCTD inhibits glioma cell growth and induces cell apoptosis,and the anticancer activity of NCTD may be induced through targeting bcl-2 anti-apoptotic family members.

关 键 词：胶质瘤 去甲斑蝥素 凋亡 B细胞淋巴瘤/白血病-2基因 

分 类 号：R739.41[医药卫生—肿瘤]