内源性保护蛋白Survivin对辐射诱导细胞凋亡的抑制作用  

Inhibitory effect of endogenous protective protein Survivin on apoptosis induced by radiation

作　　者：高洋[1] 黎海蒂[1] 罗成基[1] 高京生[1] 郑业华[1] 

机构地区：[1]解放军第三军医大学预防医学院全军复合伤研究所,创伤与复合伤国家重点实验室,重庆市400038

出　　处：《中国临床康复》2004年第35期8028-8029,i004,共3页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:抗凋亡蛋白Survivin对辐射诱导的细胞凋亡表现出很强的抑制作用。采用低氧预处理方法,诱导IEC-6细胞表达Survivin,观察其对辐射诱导的细胞凋亡的影响,并研究其可能机制,可为放射病的防护提供新的方法和实验依据。方法:分为空白组,单纯放射组,低氧+放射组和拮抗组4组。①将IEC-6细胞置于20mL/LO2环境中低氧6,8,12h,复氧24,48h,用Western-blot检测Survivin。②给予各实验组35Gy60Co照射,采用琼脂糖凝胶电泳分析DNA片段化,及采用细胞流式PI法检测细胞凋亡率。结果:低氧预处理后出现Survivin蛋白沉积带,以低氧预处理8h复氧48h组最明显。拮抗组、单纯放射组出现典型的DNA片段梯现象,低氧+放射组明显轻于上述两组。单纯放射3h凋亡率由1.44±0.08升高到8.57±0.27,6h达到凋亡高峰24.44±0.32。低氧+放射组凋亡率在3h仅为6.33±0.19,6,12,24h凋亡率明显低于单纯放射组。拮抗组凋亡率明显高于低氧+放射组。结论:低氧预处理可诱导内源性保护蛋白Survivin表达,Survivin可减轻辐射诱导的细胞DNA损伤,同时明显降低辐射诱导的细胞凋亡率。AIM:Antiapoptotic protein Survivin shows great inhibition on radiation-induced apoptosis.Survivin expression of ICE-6 is induced with hypoxic preconditioning method to observe the effect of Survivin on radiation-induced apoptosis and study the possible mechanism of it.The research may provide a new method and an experimental basis for protecting the radial diseases. METHODS:The cells were divided into four groups:control group,radiation group,hypoxia plus radiation group, and inhibition group.①IEC-6 cells were put on 20 mL/L O2 hypoxia environment for 6,8 and 12 hours and followed by reoxygenation for 24 and 48 hours,and then the expression of Survivin was detected with western-blotting.②Each group was treated with 35Gy60Co,and then the apoptosis rate and the DNA fragmentation were examined with the flow cytometry and agarose gel electrophoresis. RESULTS:Survivin protein sediment was found after hypoxia preconditioning and the group of hypoxia 8 hours and followed by reoxygenation 48 hours was the most obvious one.DNA ladder was found in the inhibition group and the radiation group,and it was significantly reduced in the hypoxia plus radiation group.The apoptosis rate of radiation for 3 hours was increased from 1.44±0.08 to 8.57±0.27.Radiation for 6 hours reached its peak(24.44±0.32).Hypoxia plus radiation for 3 hours was 6.33±0.19,and it was significantly lower than that in the radiation group at 6,12,and 24 hours.The apoptosis rate of the inhibition group was significantly higher than that of the hypoxia plus radiation group. CONCLUSION:Hypoxia preconditioning can induce the expression of Survivin and Survivin can decrease the apoptosis rate and DNA damage induced by radiation.

关 键 词：放射 辐射诱导 低氧预处理 凋亡率 细胞凋亡 蛋白S IEC-6细胞 细胞DNA DNA片段 拮抗 

分 类 号：R730.55[医药卫生—肿瘤] R364.4[医药卫生—临床医学]