辐射损伤对HRAD17基因转染不同类型细胞的影响  

Study of the effect of radiation damage on different kinds of cells transfected with HRAD17

作　　者：钱鑫[1] 韩云[1] 孙艳[1] 杨业鹏[2] 朱应葆[1] 

机构地区：[1]北京大学第三医院中心实验室细胞与分子生物学室,100083 [2]北京大学第三医院基础医学院放射医学教研室

出　　处：《中华放射医学与防护杂志》2006年第1期3-5,共3页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金资助项目(30470976)

摘　　要：目的探讨HRAD17在DNA辐射损伤修复及辐射诱导的凋亡中的作用。方法利用基因转染技术,对小鼠成纤维细胞NIH3T3和人宫颈癌HeLa细胞均分别转染pDOR-neo载体,pDOR-HRAD17s(正义)和pDOR-HRAD17a(反义)质粒,G418筛选获得阳性克隆。转染后各组细胞去血清培养48 h同步化后给予60Coγ射线10 Gy照射,吸收剂量率为2.54 Gy/min,加入完全培养基继续培养,并于0、61、2、14、36和48 h收集固定细胞,碘化丙啶(PI)避光染色后,流式细胞仪检测细胞DNA量。结果在辐射损伤后,转染正义HRAD17的成纤维细胞主要发生G1/S期阻滞,转染正义HRAD17的肿瘤细胞主要发生G2/M期阻滞;转染正义HRAD17的细胞凋亡比例降低,转染反义HRAD17的细胞凋亡比例升高。结论HRAD17同时参与细胞周期G1/S和G2/M期阻滞,并抑制辐射损伤诱导的细胞凋亡。Objective To study the role of HRAD17 in DNA damage and apoptosis induced by ionizing radiation. Methods Plasmids including pDOR-neo vector, pDOR- HRAD17s （sense） and pDOR- HRAD17a （anti-sense） were transfected into mouse fibroblasts （NIH3T3） and human uterine cervix cancer cells （HeLa）. Positive clones were screened by G418. After 48 h of serume-free culture, all kinds of transfected cells were exposed to 10 Gy γ-rays （at 2.54 Gy/min） and immediately cultured with serum. The cells were isolated and fixed by 70% ethanol, with were collected at 0, 6, 12, 24, 36 and 48 h post-irradiation, and stained by propedium iodide （PI） in the dark. Flow cytometry was performed to detect DNA in these cells. Results Ionizing radiation could induce G1/S arrest in normal cells transfected with sense HRAD17, but G2/M arrest in tumour ceils. Interestingly, cells transfected with antisense HRAD17 showed an elevated apoptotic rate compared to the control group. Conclusion HRAD17 is able to function in both G1/S and G2/M arrest and inhibit apoptosis induced by radiation damage.

关 键 词：HRAD17 辐射损伤 细胞周期 凋亡 基因转染 类型 

分 类 号：R730.5[医药卫生—肿瘤]