5cGy γ射线照射正常人淋巴母细胞基因表达转录谱的变化  

ALTERATIONS OF TRANSCRIPTIONAL PROFILE IN NORMAL HUMAN LYMPHOBLASTOID CELLS BY 5cGy OF γ-RAY EXPOSURE

作　　者：龙贤辉[1] 徐勤枝[1] 贺性鹏[2] 隋建丽[1] 安静[1] 白贝[1] 周平坤[1] 

机构地区：[1]北京放射与辐射医学研究所,北京100850 [2]南华大学公共卫生学院,湖南衡阳市421000

出　　处：《辐射防护》2006年第2期78-84,F0004,共8页Radiation Protection

基　　金：国家高技术研究发展计划(863)资助项目(编号:2004AA221160);国家自然科学基金资助项目(编号:30270423)

摘　　要：应用基因芯片技术分析5 cGy低剂量60Coγ射线照射正常人淋巴母细胞(AHH-1)后基因转录产物水平变化,探讨低剂量辐射对基因表达的影响规律。5 cGy60Coγ射线照射AHH-1细胞后4 h,提取总RNA,用包含有14 112个基因探针的人类cDNA芯片分析基因转录谱,照射组与对照组细胞的基因表达量差异比值大于2或小于0.5,两次检测结果一致的基因点为有效差异表达基因;用RT-PCR进一步验证部分差异表达基因;Western blot分析蛋白表达变化。结果筛选出了11个表达上调基因,9个表达下降基因;RT-PCR检测结果与芯片结果相一致,包括Connexin43、BMPR2、NOL6、LOC51760、LYK5等基因;Westernblot证实Connexin43基因在翻译水平表达亦增加。实验结果表明所筛选出的差异表达基因有助于阐述低剂量辐射生物效应机理,部分基因有可能成为低剂量辐射暴露的生物标志物。In order to investigate the systematic effects of low dose irradiation on transcription regulation of normal human lymphoblastoid cells, cDNA chip containing the probes related to 14 112 human genes was applied to assay the alterations of transcriptional profile of AHH-1 cells exposed to 5 cGy of γ-ray from 60^Co. After a repeat genechip hybridization the genes with 2-fold increase or decrease in expression signal is regarded as a differential expression gene. Then RT-PCR and Western blot were used to confirm the results of genechips. The results demonstrated that 11 genes were up-regulated and 9 genes were down-regulated in AHH-1 cells at 4 h post to 5 cGy γ-ray irradiation. The changed transcriptional level was further confirmed by RT-PCR assay for some of these genes, i. e. Connexin43, BMPR2, NOL6, LOC5176, LYK5, etc. Up-regulation of protein expression of Connexin43 was also Connexin43 by Western blot assay. The observation of these differential expressed genes induced by 5 cGy γ-my will help to understand the mechanism of biological effects of low dose irradiation. New biomarkers of low dose exposure might be developed from these genes.

关 键 词：基因芯片 基因表达 电离辐射 细胞周期调控 

分 类 号：R811.5[医药卫生—放射医学]