运用抑制消减杂交构建辐射诱导EST库  

Suppression subtractive hybridization in construction of radiation-induced EST library

作　　者：铁轶[1] 罗瑛[1] 隋建丽[1] 周平坤[1] 

机构地区：[1]北京放射医学研究所,100850

出　　处：《中华放射医学与防护杂志》2001年第1期9-11,共3页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金!资助项目 (39870 2 14 );全军医学卫生科研基金!资助项目 (990 5 0 0 8)

摘　　要：目的　克隆并鉴定低剂量辐射诱导基因 ,为探讨低剂量辐射生物学效应分子机理提供有益信息。方法　 0 5Gyγ射线照射人胚肺细胞后 ,用抑制消减杂交技术 (SuppressionSubtractiveHybridization ,SSH)构建低剂量辐射诱导EST库。用反向Northern杂交法对库中的EST片段进行筛选 ,挑取阳性克隆测序 ,并在GenBank序列数据库中进行同源性比较。结果　得到受照后表达升高 90个EST序列 ,代表 2 1个基因的转录产物 ,部分基因的功能已明确。从功能上分析 ,这些基因产物的生物学作用涵盖了细胞周期、信号转导、细胞骨架、代谢、蛋白合成等几个重要的细胞生物学代谢反应过程 ,显示出细胞对低剂量辐射反应过程的多样性。另外 ,实验还获得 4个新cDNA序列。结论　利用抑制消减杂交技术 ,从辐射前后HEL细胞中 ,得到低剂量辐射诱导EST片段 ,它们与细胞周期、增殖、代谢、信号转导、应激反应等相关 ,这些基因可能在细胞的低剂量辐射效应中起到重要作用。Objective\ To clone and identify radiation\|induced genes from 0 5 Gy γ ray irradiated human embryo lung cells(HEL).The identification and functional studies of radiation\|induced genes will prompt the elucidation of the molecular mechanisms of low dose radiation\|induced biological effects.It will also profit the understanding of the basic processes of cellular metabolisms. Methods\ A low dose radiation\|induced differentially displaying EST library has been constructed by suppression subtractive hybridization from HEL cells irradiated with 0 5 Gy γ rays.The EST library was screened by reverse Northern hybridization analysis.Positive clones were sequenced and the similarity was searched against the DNA database in GenBank. Results\ Altogether 90 positive cDNA clones with increased expression after 0 5 Gy irradiation were identified which corresponded to 21 individual genes.These genes involved in the processes of cell cycle control,signal transduction,cell skeleton,metabolism and protein synthesis,etc.All these demonstrated the diverse responses of cells to low dose radiation.Moreover,four novel cDNA were obtained. Conclusions\ Low dose radiation induces ESTs which relate to cell proliferation,cell cycle control,signal transduction,cell skeleton,metabolism,protein synthesis and stress response etc were cloned and identified by SSH.Our data suggest that these genes could play important roles in the biological response of cells to low dose radiation.\;

关 键 词：辐射诱导基因 抑制消减杂交 低剂量 差异显示EST库 

分 类 号：R312[医药卫生—基础医学]