CAJ | 학술논문

目的探讨Bcl-XL小分子干扰RNA(siRNA)在逆转人结肠癌细胞获得性肿瘤坏死因子相关的凋亡诱导配体(TRAIL)耐药中的作用.方法以Bcl-XL siRNA转染获得性TRAIL耐药的人结肠癌DLD1-TRAIL/R细胞24 h,并用TRAIL蛋白处理,通过细胞计数和流式细胞仪检测细胞的存活率,并通过Western blot法检测各种凋亡相关蛋白的活化情况.结果Bcl-XL siRNA能有效下调DLD1-TRAIL/R细胞中Bcl-XL蛋白的表达水平,并且能够有效地逆转其对TRAIL蛋白的耐药.DLD1-TRAIL/R细胞经过Bcl-XL siRNA和TRAIL蛋白共同处理2 h后,其细胞凋亡率＞50%;共同处理4 h后,其细胞存活率＜40%;而对照组和TRAIL蛋白处理组的细胞凋亡率和生存率无明显变化(P＜0·05).Western blot法检测结果表明,经Bcl-XL siRNA与TRAIL蛋白联合处理后,DLD1TRAIL/R细胞中caspase-8、caspase-9、Bid、caspase-3以及多聚腺苷酸二磷酸核糖聚合酶(PARP)均明显活化,细胞色素C的释放显著增加.结论 Bcl-XL siRNA能够有效逆转结肠癌细胞获得性TRAIL耐药,这将为治疗肿瘤耐药提供一种新的思路.
목적 탐토Bcl-XL소분자간우RNA(siRNA)재역전인결장암세포획득성종류배사인자상관적조망유도배체(TRAIL)내약중적작용.방법 이Bcl-XL siRNA전염획득성TRAIL내약적인결장암DLD1-TRAIL/R세포24 h,병용TRAIL단백처리,통과세포계수화류식세포의검측세포적존활솔,병통과Western blot법검측각충조망상관단백적활화정황.결과Bcl-XL siRNA능유효하조DLD1-TRAIL/R세포중Bcl-XL단백적표체수평,병차능구유효지역전기대TRAIL단백적내약.DLD1-TRAIL/R세포경과Bcl-XL siRNA화TRAIL단백공동처리2 h후,기세포조망솔＞50%;공동처리4 h후,기세포존활솔＜40%;이대조조화TRAIL단백처리조적세포조망솔화생존솔무명현변화(P＜0·05).Western blot법검측결과표명,경Bcl-XL siRNA여TRAIL단백연합처리후,DLD1TRAIL/R세포중caspase-8、caspase-9、Bid、caspase-3이급다취선감산이린산핵당취합매(PARP)균명현활화,세포색소C적석방현저증가.결론 Bcl-XL siRNA능구유효역전결장암세포획득성TRAIL내약,저장위치료종류내약제공일충신적사로.
Objective To investigate the reversing effct of Bcl-XL small interfering RNA(siRNA)on the acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) in human colon cancer.Methods Human colon cancer cells DLDl.TRAIL/R,with acquired resistance to TRAIL,were firstly transfected with Bcl-XL siRNA for 24 h followed by the treatment of TRAIL protein.The survival rate of DLD1-TRAIL/R cells was assessed by FACS analysis and cell number counting,respectively,and activation of its apoptotic signaling was evaluated by Western blot.Results Bcl-XL siRNA effectivelv down regulated the expression of Bcl-XL protein and reversed the acquired resistance to TRAIL in DLD1-TRAIL/R cells-After combination treatment of Bcl-XL siRNA and TRAIL protein.the apoptotic rate of DLD1-TRAIL/R eells was more than 50%and survival rate was less than 40%,whereas there was no efiect on the survival of DLD1-TRAIL/R cells after treatment with control treatment or TRAIL protein treatment alone ( P＜0.05).Western blot analysis demonstrated that caspase-8,caspase-9,Bid,caspase-3,and p01y(ADPribose)polymerase(PARP)were obviously activated after combination treatment with Bcl-XL siRNA and TRAIL protein,and the release of cytochrome C was also significantly increased.Conclusion Bcl-XLsiRNA can effectively reverse the acquired resistance to TRAIL in human colon cancer cells,suggesting that it might be a new strategy for overcoming the resistance in cancer therapy.