CAJ | 학술논문

目的于体外实验观察人重组肿瘤坏死因子相关凋亡诱导配体(rhTRAIL)联合环氧化酶-2选择性抑制剂塞来昔布对胆囊癌的疗效,初步探讨产生疗效的机制.方法用western-blot法检测塞来昔布作用于胆囊癌细胞株后,抗凋亡蛋白c-FLIP和死亡受体DR4、DR5表达状况.rhTRAIL联合塞来昔布作用于胆囊癌细胞株SGC-996后,采用3种方法检测细胞凋亡状况:(1)细胞的相差显微镜观察;(2)caspase 3、7活性检测;(3)Annexin染色后凋亡细胞流式细胞仪检测.结果塞来昔布可下调SGC-996细胞c-FLIPs的表达,上调DR5的表达,且呈浓度、时间依赖性.rhTRAIL联合塞来昔布作用于胆囊癌细胞后,细胞凋亡水平明显高于单独用药组和对照组.结论塞来昔布可通过下调c-FLIPs表达和上调DR5表达,显著增强rhTRAIL诱导胆囊癌SGC-996细胞凋亡的作用.
목적우체외실험관찰인중조종류배사인자상관조망유도배체(rhTRAIL)연합배양화매-2선택성억제제새래석포대담낭암적료효,초보탐토산생료효적궤제.방법용western-blot법검측새래석포작용우담낭암세포주후,항조망단백c-FLIP화사망수체DR4、DR5표체상황.rhTRAIL연합새래석포작용우담낭암세포주SGC-996후,채용3충방법검측세포조망상황:(1)세포적상차현미경관찰;(2)caspase 3、7활성검측;(3)Annexin염색후조망세포류식세포의검측.결과새래석포가하조SGC-996세포c-FLIPs적표체,상조DR5적표체,차정농도、시간의뢰성.rhTRAIL연합새래석포작용우담낭암세포후,세포조망수평명현고우단독용약조화대조조.결론새래석포가통과하조c-FLIPs표체화상조DR5표체,현저증강rhTRAIL유도담낭암SGC-996세포조망적작용.
Objective To observe the effect of combined treatment with rhTRAIL(recombinant human TNF-related apoptosis inducing ligand) and selective Cox-2 inhibitor Celecoxib on gallbladder carcinoma in vitro and to explore the possible mechanism of the effect. Methods Western blot analysis was used to detect the expression of c-FLIP and death receptors after treatment by Celecoxib. Apoptosis of gallbladder cell line SGC-996 after the combined treatment with Celecoxib and rhTRAIL was detected in three ways: (1) phase microscopy of the cells, (2) detection of effector caspase-3 and caspase-7 activity, and (3) determination of the proportion of apoptotic cells labeled by Annexin V-PI flow cytometric analysis using CELLQUEST software. Results Celecoxib down-regulated the expression of c-FLIPs and up-regulated the expression of DR5 in a dose- and time-dependent mode on cell line SGC-996. Apoptotic levels in the combined treatment group in cell line SGC-996 were significantly higher than those in the single drug treatment group and control group. Conclusion Celecoxib markedly sensitized rhTRAIL-induced apoptosis through the down-regulation of c-FLIPs and up-regulation of DR5 in gallbladder carcinoma cell line SGC-996.