Smac基因联合^12C^6＋离子照射对膀胱癌EJ细胞的生物效应观察  

Observation on bio-effect of EJ cell line with smac gene transferred and ^12C^6＋ ions-ray irradiation

作　　者：赵宝锋[1] 田梅[1] 阮健磊[1] 苏旭[1] 李文建[2] 

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所, 北京100088 [2]中国科学院近代物理研究所

出　　处：《中华放射医学与防护杂志》2007年第1期67-69,共3页Chinese Journal of Radiological Medicine and Protection

基　　金：中国博士后基金资助项目（2005038364）;兰州重离子加速器国家实验室基金资助项目

摘　　要：目的观察smac基因能否提高膀胱癌EK细胞^12C^6＋离子照射的生物效应。方法实验分为3组：空白对照组、转染空载体组、转染smac基因组。利用免疫组化及流式细胞仪方法检测3组细胞内smac基因表达。转染24h后对3组分别进行0、2和4Gy ^12C^6＋离子照射，用克隆形成实验对3个组分别检测细胞的存活分数并利用流式细胞仪检测细胞凋亡率。结果免疫组化结果表明转染pcDNA3．1／smac的细胞其smac基因表达量明显高于转染空载体pcDNA3．1的细胞和空白组细胞。流式细胞仪数据显示，smac基因转染细胞smac基因的荧光强度（3080）显著高于转染空载体组（2256）（P〈0．05），而后者与空白对照组间比较差异无统计学意义（P〉0．05）。克隆形成实验结果表明，转染smac基因组细胞^12C^6＋离子照射后的存活分数较转染空载体组细胞显著降低（P〈0．05）。流式细胞仪测定结果表明，^12C^6＋离子照射的转染smac基因组的细胞凋亡率明显高于转染空载体组（P〈0．05）。结论外源smac基因转染膀胱癌细胞株能显示高表达，并能提高膀胱癌细胞的凋亡率，与^12C^6＋离子照射结合后能显著降低细胞存活分数和进一步增高凋亡率。Objective To observe whether smac gene can increase the bio-effect of EJ cells induced by ^12C^6＋ ions irradiation. Methods The cells were divided into three groups- blank control group, vector transferred group and smac gene transferred group. Immuno-histochemistry and flow cytometry were used to identify the expression of smac gene. Following ^12C^6＋ ions treatment at 0, 2, 4 Gy, three groups＇ survival ratio was detected by colony-formation assay. 24 h after ^12C^6＋ ions treatment, flow cytometry was used to detect apoptosis. Results Immuno-histochemistry resuhs indicated that the expression of smac gene in smac gene transferred group was notably enhanced compared with vector pcDNA3.1 transferred group. Fluorescence- activated cell sorting （FACS） resuhs also indicated that the expression of smac gene in smac gene transferred group was significantly enhanced compared with that of vector pcDNA3.1 transferred group （3080 vs 2256, P 〈 0.05）, but there is no significant difference between vector pcDNA3.1 transferred group and blank control group. The results of colony-formation assay suggested that the smac gene transferred group showed significantly lower survival fraction than vector pcDNA3.1 transferred group （P 〈 0.05）. 24 h after ^12C^6＋ ions treatment, FACS results indicated that smac gene transferred group showed apparent cell apoptosis ratio, which was higher than that of vector peDNA3.1 transferred group （ P 〈 0.05）. Conclusions Smac gene could show high expression in smae gene transferred group cells and increase the apoptosis ratio of EJ cells. Smac gene could further enhance the apoptosis and notably decrease the survival fraction of EJ cells when combined with ^12C^6＋ ions irradiation.

关 键 词：SMAC基因 碳离子 放射治疗 EJ细胞 细胞凋亡 

分 类 号：R686[医药卫生—骨科学]