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戴德坚; 陆才德; 郭俊明; 章军
癌症, 2005, Volume: 24, Issue: 8Journal Article
R735.7; 背景与目的:由于Survivin在大多数肿瘤细胞中过表达,而在正常成人终末分化组织中沉默,并参与了恶性肿瘤的发生、发展和化疗耐药,使其成为值得关注的抗癌治疗靶.反义寡核苷酸可用来封闭凋亡抑制基因Survivin,诱导肿瘤细胞凋亡或增强其化疗敏感性.本研究旨在探讨反义抑制Survivin表达对肝癌细胞系阿霉素敏感性的影响.方法:RT-PCR、Western blot检测HepG2和HepG2/ADM细胞Survivin表达;MTF法评估HepG2和HepG2/ADM对反义复合物和阿霉素的敏感性;不同浓度的反义复合物分别转染HepG2和HepG2/ADM细胞,RT-PCR检测SurvivinmRNA表达;等辐射分析法评估不同浓度反义复合物和亚致死浓度阿霉素对HepG2和HepG2/ADM细胞的协同效应:流式细胞术检测激活型Caspase-3表达及凋亡率.结果:HepG2/ADM细胞Survivin mRNA是HepG2细胞的15倍,蛋白水平是HepG2的18倍.两种细胞均显示对反义复合物敏感并呈浓度依赖关系,反义复合物对HepG2和HepG2/ADM细胞的IC50分别为317.90 nmol/L和480.74 nmol/L,在500 nmol/L时抑制率达到71.10%和53.67%.ADM对HepG2和HepG2/ADM细胞的IC50分别为0.36 μg/ml和2.12 μg/ml,耐药指数为6.反义复合物剂量依赖性下调HepG2和HepG2/ADM细胞Survivin mRNA表达,对HepG2和HepG2/ADM细胞的IC50分别为271.93 nmol/和365.72nmol/L,400 nmol/L时表达水平分别下调了69.12%和60.01%.反义复合物和低浓度ADM联合使HepG2细胞的敏感性增加了6倍,使HepG2/ADM细胞的敏感性增加了4倍.联合处理激活了HepG2/ADM细胞Caspase-3活性,诱导细胞凋亡,激活型Caspase-3和凋亡率随反义复合物浓度的增加而逐渐递增.结论:反义抑制Survivin表达增强肝癌细胞系对阿霉素的敏感性,反义寡核苷酸和阿霉素联合可能是耐阿霉素肝细胞癌临床治疗的一种合理策略.
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