二硫代氨基甲酸吡咯烷对人骨肉瘤细胞凋亡及黑色素瘤凋亡抑制蛋白和半胱氨酰天冬氨酸特异性蛋白酶-3表达的影响  

Effects of pyrrolidine dithiocarbamate on apoptosis and expression of Livin and cysteinyl aspartate-specific protease-3 in human osteosarcoma cells

作　　者：王玉峰[1] 黄涛[1] 叶天宝 成硕[1] 胡春吉[1] 柴利鹏[1] Wang Yufeng Huang Tao Ye Tianbao Cheng Shuo Hu Chunji Chai Lipeng(Department of Orthopaedics, the First Affiliated Hospital of China Medical University, Shenyang 110001, China)

机构地区：[1]中国医科大学附属第一医院骨科,沈阳110001

出　　处：《中华实验外科杂志》2016年第11期2496-2498,共3页Chinese Journal of Experimental Surgery

基　　金：辽宁省科学技术计划项目（2012225071）

摘　　要：目的 观察二硫代氨基甲酸吡咯烷对人骨肉瘤MG-63细胞凋亡及黑色素瘤凋亡抑制蛋白（Livin）和半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3表达的影响.方法 采用体外培养人骨肉瘤MG-63细胞株,用50、100、200、400 μmol/L的吡咯烷二硫代氨基甲酸盐（PDTC）分别作用于骨肉瘤MG-63细胞,在24、48、72 h后,噻唑蓝（MTT）法测各组细胞存活率,用流式细胞仪检测细胞的凋亡率.作用于骨肉瘤MG-63细胞24h后,用Western blot方法检测各组细胞Livin和Caspase-3蛋白的表达水平.结果 随PDTC浓度增加骨肉瘤MG-63细胞增殖活性呈下降趋势,作用时间越长细胞的增殖活性越低,而细胞的凋亡率呈上升趋势（P＜0.05）.作用24h后,随PDTC浓度增加各组细胞中Livin蛋白的表达为1.384±0.009、0.912±0.007、1.501±0.011、0.751 ±0.008,对照组为1.834±0.006,呈下降趋势,而Caspase-3蛋白表达呈上升趋势（P＜0.05）.结论 PDTC可诱导入骨肉瘤MG-63细胞凋亡,其机制可能与PDTC抑制核因子（NF）-κB传导通路,下调Livin表达继而上调Caspase-3表达有关.Objective To detect the effect of pyrrolidine dithiocarbamate （PDTC） on apoptosis and expression of Livin and cysteinyl aspartate-specific protease-3 （Caspase-3） in human osteosarcoma cell MG-63.Methods MG-63 cells were cultured in vitro.At 24,48 and 72 h after treatment with PDTC at different concentrations （50,100,200 and 400μmol/L）,methyl thiazol tetrazolium （MTT） assay was used to observe the growth inhibition of MG-63 cells.The cell apoptosis was assessed by flow cytometry.Protein of Livin and Caspase-3 was detected by Western blotting at 48 h after treatment with PDTC at different concentrations.Results PDTC could inhibit growth and increase apoptosis of MG-63 cells in a dose-and time-dependent manner.PDTC also down-regulated the protein expression of Livin.After treatment with PDTC for 24 h,the protein expression of Livin was 1.384 ± 0.009,0.912 ± 0.007,1.501 ±0.011,and 0.751 ±0.008,and that in the control group was 1.834 ±0.006 （P〈0.05）,and the protein expression of Caspase-3 was elevated （P 〈 0.05）.Conclusion PDTC can induce apoptosis of MG-63 cells,which may be correlated with down-regulation of Livin and up-regulation of Caspase-3 expression.

关 键 词：二硫代氨基甲酸吡咯烷 骨肉瘤 黑色素瘤凋亡抑制蛋白 半胱氨酰天冬氨酸特异性蛋白酶-3 Cysteinyl aspartate-specific protease-3 

分 类 号：R285.5[医药卫生—中药学]