Bcl-XL小分子干扰RNA逆转人结肠癌获得性肿瘤坏死因子相关凋亡诱导配体的耐药  

Overcoming acquired resistance to tumor necrosis factor-related apoptosis-inducing Iigand by Bcl-XLsmall interfering RNA in human colon cancer

作　　者：朱洪波[1] 黄学锋[1] 胡静姿[2] 周玮[1] 陈薇[1] 陈琳琳[1] 何超[1] 

机构地区：[1]浙江大学医学院附属邵逸夫医院肛肠外科生物医学研究中心,杭州310016 [2]南京军区杭州疗养院空勤疗养区

出　　处：《中华肿瘤杂志》2008年第4期245-249,共5页Chinese Journal of Oncology

基　　金：国家自然科学基金资助项目（30528030、30572153）;浙江省自然科学基金资助项目（Y205093）

摘　　要：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24h，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。DLDl-TRAIL／R细胞经过Bcl-XLsiRNA和TRAIL蛋白共同处理2h后，其细胞凋亡率〉50％；共同处理4h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，DLDl-TRAIL／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。Objective To investigate the reversing effect of Bel-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 DLD1-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 effectively downregulated 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 cells was more than 50% and survival rate was less than 40%, whereas there was no effect 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 poly （ADP- ribose） 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-XL siRNA 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.

关 键 词：BCL-XL 小分子干扰RNA 肿瘤坏死因子相关的凋亡诱导配体 耐药 

分 类 号：R686[医药卫生—骨科学]