中华放射医学与防护杂志

钱鑫,韩云,孙艳,等.辐射损伤对HRAD17基因转染不同类型细胞的影响[J].中华放射医学与防护杂志,2006,26(1):3-5.QIAN Xin,HAN Yun,SUN Yan,et al.Study of the effect of radiation damage on different kinds of cells transfected with HRAD17[J].Chin J Radiol Med Prot,2006,26(1):3-5

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

投稿时间：2005-09-28

DOI：

中文关键词: HRAD17 辐射损伤细胞周期凋亡

英文关键词:HRAD17 Radiation damage Cell cycle Apoptosis

基金项目:国家自然科学基金资助项目(30470976)

作者	单位	E-mail
钱鑫	100083 北京大学第三医院中心实验室细胞与分子生物学室	yingbaozhu@medmail.com.cn
韩云	100083 北京大学第三医院中心实验室细胞与分子生物学室
孙艳	100083 北京大学第三医院中心实验室细胞与分子生物学室
杨业鹏	基础医学院放射医学教研室
朱应葆	100083 北京大学第三医院中心实验室细胞与分子生物学室

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中文摘要:

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48h同步化后给予⁶⁰Coγ射线10Gy照射,吸收剂量率为2.54Gy/min,加入完全培养基继续培养,并于0、61、2、14、36和48h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。

英文摘要:

Objective To study the role of HRAD17 in DNA damage and apoptosis induced by ionizing radiation. Methods Plasmids including pDOR-neo vector,pDORHRAD17s（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 cells.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.

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