小干扰RNA沉默Bcl-2表达增强食管癌细胞放射敏感性研究  

Small interfering RNA in silencing Bcl-2 expression and enhancing radiosensitivity of esophageal cancer cells

作　　者：刘冬梅[1] 李国文[1] 田希凤[1] 张松辉[1] 

机构地区：[1]河南省高等学校临床医学重点学科开放实验室郑州大学基础医学院放射医学专业郑州大学第一附属医院放疗科,郑州450052

出　　处：《中华放射肿瘤学杂志》2011年第5期435-437,共3页Chinese Journal of Radiation Oncology

摘　　要：目的探讨Bcl-2基因的特异性小干扰RNA（siRNA）对食管癌细胞（EC9706细胞）Bcl-2蛋白表达和凋亡的影响及放射增敏作用。方法化学合成Bcl-2基因的siRNA，通过脂质体转染法转入EC9706细胞。流式细胞仪检测转染前后Bcl-2蛋白表达及细胞凋亡，成克隆分析siRNA联合x线照射对EC9706细胞的放射敏感性影响。结果Bcl-2siRNA2、Bcl-2siRNA2、Bcl-2siRNA，转染后Bcl-2蛋白在EC9706细胞中的阳性表达率分别为25．13％、38．87％、30．55％，较对照组的84．28％明显下降（t=4．01、3．04、3．64，P均〈0．05）。Bcl-2siRNA．转染组凋亡率为33．86％，明显高于空白对照组的5．51％（t=6．55，P〈0．01）和阴性siRNA1转染组的5．59％（t=6．54，P〈0．01）；Bcl-2siRNA1联合x线照射的细胞凋亡率为56．76％，明显高于单纯照射组的24．51％（t=3．59，P〈0．05）。成克隆分析的Bcl-2siRNA，转染加照射组D0、Dq、SF2值均低于单纯照射组，放射增敏比为1．33（D0值之比）。结论Bcl-2基因的特异性siRNA可明显增强EC9706细胞的放射敏感性，具有良好临床应用前景。Objective To explore the effects of small interfering RNA （siRNA） specific to Bcl-2 gene on radiosensitivity of esophageal cancer cells. Methods Bcl-2 gene siRNA （ Bcl-2 siRNA ） was induced into esophageal cancer EC9706 cells by lipofectamine. Bcl-2 protein expression and apoptosis of EC9706 cells were detected by flowcytometer. Clone forming assay was used to determine the inhibitory effects of X-ray radiation combined with Bcl-2 siRNA interference. Results When Bcl-2 siRNA had been induced into EC9706 cells, Bcl-2 protein expression in EC9706 cells was inhibited, and cell apoptosis was increased. Bcl-2 protein expression rates of EC9706 cells induced with Bcl-2 siRNA1 , A2, A3 （25.13% ± 2. 04% ,38.87% ± 3.34%, 30.55% ± 2. 73% ） were lower than the control group （84.28% ± 1.47% ） （t =4. 01,3.04,3.64, P 〈 0. 05 ）. After interference, the apoptosis rate of EC9706 cells （ 33.86% ±1.04% ） was higher than the control group and siRNA negative group （5.51% ± 0. 14% and 5.59% ±0.46% ）（ t =6. 55,6. 54 ,P 〈 0. 01 ）. Bcl-2 gene siRNA interference enhanced X-ray inducing apoptosis of EC9706 cells （56.76% ± 1.24% ）, which was higher than the radiation alone group （24.51% ±0.48% ） （t = 3.59,P 〈0. 05 ）. The D0, Dq, and SF2 of combined treatment group were much lower than those of irradiation alone group . The sensitization enhancing ratio was 1.32 （ ratio of DO values ） . Conclusions Bcl-2 gene siRNA could enhance the radiosensitivity of esophageal cancer EC9706 cells and may has a good future in clinical practice.

关 键 词：核糖核酸干扰 放射敏感性 BCL-2表达 细胞系 食管肿瘤 

分 类 号：R735[医药卫生—肿瘤]