低剂量γ射线辐照对人淋巴母细胞基因表达转录谱的影响  

Effects of low dose γ-ray on transcription profiles of human lymphoblastoid cells

作　　者：何颖[1] 沈先荣[1] 钱甜甜[1] 王庆蓉[1] 蒋定文[1] 刘玉明[1] 侯登勇[1] 陈伟[1] 李珂娴[1] 刘琼[1] 

机构地区：[1]海军医学研究所,上海200433

出　　处：《辐射研究与辐射工艺学报》2013年第5期20-29,共10页Journal of Radiation Research and Radiation Processing

基　　金：全军计划科研项目(AHJ09J012);总后"十一五"科技攻关项目(2009183006)资助

摘　　要：为探讨低剂量电离辐射生物效应作用机制,本文研究了不同剂量单次辐照后,人淋巴母细胞基因表达转录谱的变化。采用0.1 Gy、0.5 Gy和1.0 Gy剂量的γ射线照射人淋巴母细胞,未照射细胞为对照组。照射后4 h提取细胞总RNA,用含有45 033个基因的人类全基因组表达谱芯片检测分析。对差异表达基因进行层次聚类分析、基因本体论分析和通路分析,并用实时荧光定量PCR验证。结果表明,3个剂量组均上调表达的基因1330个,下调表达的基因1002个。基因表达量与吸收剂量相关的基因共18个,其中与剂量正相关的基因16个,负相关的基因2个。层次聚类分析结果表明,4个实验组中,0.1 Gy和1.0 Gy照射组差异表达基因相似程度最高。这些差异表达的基因涉及到多条通路,如细胞周期、p53信号通路、碱基切除修复、RNA转运和内质网蛋白加工等。实时荧光定量PCR检测结果表明,CASP9(caspase 9)mRNA在照射后4 h随吸收剂量表达变化与基因芯片检测结果一致。基因芯片筛选出的差异表达基因有助于阐明低剂量辐射生物效应作用机理,与辐射剂量相关的差异表达基因有可能成为低剂量辐射暴露的生物剂量计。ABSTRACT To investigate the mechanism of biological effects induced by low dose radiation, the alterations of transcriptional profiles of human normal lymphoblastoid cell AHH-1 lines irradiated with different doses of γ-rays were assayed. AHH-1 lines were irradiated with γ-rays of 0.1, 0.5, and 1.0 Gy, respectively, with the unirradiated cells as control group. Total RNA of irradiated cells was extracted 4 h after irradiation, and then cDNA microarray was used to detect the transcriptional profiles of the ceils. The different expressions of genes were assayed by hierarchical cluster analysis, gene ontology and pathway analysis, and then identified by real-time Q-PCR. The results of real-time Q-PCR of CASP9 were in accordance with microarray assays. The microarray assays showed that there were 1 330 up-regulated genes and 1 002 down-regulated genes in three irradiated groups. Alterations of 18 genes expression were related with absorbed dose. Among these genes, 16 genes were positive relation, and 2 genes were negative relation with dose. The results of hierarchial cluster analysis indicated that the similarity of the expression profiles of 0.1 Gy and 1.0 Gy was the highest among 4 groups. These altered genes could be related to multiple signal transductions, such as cell cycle, p53 signal pathway, base excision repair, RNA transport, endoplasmic reticulum proteins processing pathways, and so on. These differential expressed genes detected by cDNA microarray would help to elucidate the mechanisms of biological effects of low dose irradiation. New biological dosimeter for low dose exposure might be developed from these dose-related genes.

关 键 词：基因芯片 低剂量电离辐射 人淋巴母细胞 基因本体论 通路分析 层次聚类分析 

分 类 号：R811.5[医药卫生—放射医学]