肿瘤坏死因子相关凋亡诱导配体联合雷公藤甲素诱导胰腺癌细胞凋亡的机制研究  

Mechanisms of tumor necrosis factor-related apoptosis inducing ligandcombined with Triptolide in inducing the apoptosis of pancreatic cancer cells

作　　者：江艳[1] 刘炜[1] 陈志宇[1] 

机构地区：[1]第三军医大学西南医院全军肝胆外科研究所 中国人民解放军西南肝胆外科医院,重庆400038

出　　处：《中华消化外科杂志》2015年第4期321-328,共8页Chinese Journal of Digestive Surgery

基　　金：国家自然科学基金（81472775）

摘　　要：目的 探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）联合雷公藤甲素（Triptolide）诱导胰腺癌细胞凋亡的机制.方法 （1）将人胰腺癌细胞MiaPaca-2（简称MiaPaca-2细胞）分为4组：空白对照组（不加任何药物）,TRAIL^＋ Triptolide^-组（仅加TRAIL）,TRAIL^-Triptolide^＋组（仅加Triptolide）和TRAIL^＋Triptolide^＋组（加TRAIL和Triptolide）.采用CCK-8检测各组细胞活力.采用Western blot检测各组细胞凋亡相关蛋白多聚ADP核糖聚合酶（PARP）、含半胱氨酸的天冬氨酸蛋白水解酶-3（Caspase-3）和含半胱氨酸的天冬氨酸蛋白水解酶-8（Caspase-8）的表达.加入Caspase-8特异性抑制剂Z-IETD-FMK后,采用CCK-8检测细胞活力,采用Caspase-Glo荧光检测法检测Caspase-8活性.采用Western blot检测各组细胞髓样细胞白血病-1（Mcl-1）、额外的大B细胞淋巴瘤（Bcl-xL）和B淋巴细胞瘤-2（Bcl-2）蛋白的表达.（2）将MiaPaca-2细胞分为8组：①TRAIL-Mcl-1 siRNA-组（不加TRAIL、不转染Mcl-1 siRNA）,TRAIL^＋ Mcl-1siRNA-组（加TRAIL、不转染Mcl-1 siRNA）,TRAIL^-Mcl-1 siRNA^＋组（不加TRAIL、转染Mcl-1 siRNA）和TRAIL^＋ Mcl-1 siRNA＋组（加TRAIL、转染Mcl-1 siRNA）.②TRAIL^-Bcl-xL siRNA-组（不加TRAIL、不转染Bcl-xL siRNA）,TRAIL^＋ Bcl-xL siRNA^-组（加TRAIL、不转染Bcl-xL siRNA）,TRAIL^-Bcl-xL siRNA^＋组（不加TRAIL、转染Bcl-xL siRNA）,TRAIL^＋ Bcl-xL siRNA^＋组（加TRAIL、转染Bcl-xL siRNA）.采用CCK-8检测各组细胞活力.采用Western blot检测各组细胞Caspase-3和Caspase-8蛋白的表达.符合正态分布的计量资料以x^-±s表示,多组间比较采用单因素方差分析,两两比较采用LSD-t检验.结果 （1）空白对照组、TRAIL^＋ Triptolide^-组、TRAIL^-Triptolide^＋组和TRAIL^＋ Triptolide^＋组MiaPaca-2细胞活力分别为100.0％±1.1％、81.2％±2.3％、78.6％±3.6％、40.1％±2.5％;PARP蛋白相对表达量分别为0.510 ±0.028、0.720±0.072、1.250±0.023、Objective To investigate the mechanisms of tumor necrosis factor-related apoptosis inducing ligand （TRAIL） combined with Triptolide in inducing the apoptosis of pancreatic cancer cells.Methods （1） The pancreatic cancer cells （MiaPaca-2 cells） were divided into 4 groups：blank control group （no drugs were added）,TRAIL^＋ Triptolide^-group （only TRAIL was added）,TRAIL^-Triptolide^＋ group （only Triptolide was added） and TRAIL^＋ Triptolide^＋ group （TRAIL and Triptolide were added）.The vitality of cells in all the 4 groups was assessed by CCK-8.The expressions of poly ADP-ribose polymerase （PARP）,cysteinyl aspartate specific proteinase-3 （Caspase-3） and Caspase-8 were detected by Western blot.The vitality of cells was detected by CCK-8 and the vitality of Caspase-8 was detected by Caspase-Glo assays after adding Z-IETD-FMK,a specific inhibitor of Caspase-8.The expressions of myeloid cell leukemia-1 （Mcl-1）,Bcl-xL and Bcl-2 were detected by Western blot.（2） The MiaPaca-2 cells were divided into 8 groups：①TRAIL^-Mcl-1 siRNA^-group （no TRAIL was added and Mcl-1 siRNA cells were not transfected）,TRAIL^＋ Mcl-1 siRNA^-group （TRAIL was added and Mcl-1 siRNA cells were not transfected）,TRAIL^-Mcl-1 siRNA^＋ group （TRAIL was not added and Mcl-1 siRNA cells were transfected）and TRAIL^＋ Mcl-1 siRNA^＋ group （TRAIL was added and Mcl-1 siRNA cells were transfected）.②TRAIL^-Bcl-xL siRNA^-group （TRAIL was not added and Bcl-xL siRNA was not transfected）,TRAIL^＋ Bcl-xL siRNA^-group （TRAIL was added and Bcl-xL siRNA was not transfected）,TRAIL^-Bcl-xL siRNA^＋ group （TRAIL was not added and Bcl-xL siRNA was transfected） and TRAIL^＋ Bcl-xL siRNA^＋ group （TRAIL was added and Bcl-xL siRNA was transfected）.The vitality of the cells in all the groups was detected by CCK-8.The expressions of Caspase-3 and Caspase-8 protein were detected by Western blot.The measurement data with normal distribution were presented as x^- ± s.The comparison am

关 键 词：胰腺肿瘤 肿瘤坏死因子相关凋亡诱导配体 雷公藤甲素 

分 类 号：R735.9[医药卫生—肿瘤]