高荣莲,陈肖华,饶亚岚,等.寡聚核苷酸芯片技术筛选电离辐射后大鼠PC12细胞的差异表达基因[J].中华放射医学与防护杂志,2005,25(3):242-245.GAO Rong-lian,CHEN Xiao-hua,RAO Ya-lan,et al.Screening for differentially expressed genes in IR-induced PC12 cells by oligonucleotide microarray[J].Chin J Radiol Med Prot,2005,25(3):242-245
寡聚核苷酸芯片技术筛选电离辐射后大鼠PC12细胞的差异表达基因
Screening for differentially expressed genes in IR-induced PC12 cells by oligonucleotide microarray
投稿时间:2005-01-04  
DOI:
中文关键词:  基因芯片  电离辐射  基因表达谱
英文关键词:Microarray  Ionizing radiation  Gene expression
基金项目:
作者单位E-mail
高荣莲 100850, 北京放射医学研究所 chenxh@nic.bmi.ac.cn 
陈肖华 100850, 北京放射医学研究所  
饶亚岚 100850, 北京放射医学研究所  
张军权 100850, 北京放射医学研究所  
王治东 100850, 北京放射医学研究所  
董波 100850, 北京放射医学研究所  
侯丽莉 100850, 北京放射医学研究所  
张浩 100850, 北京放射医学研究所  
毛秉智 100850, 北京放射医学研究所  
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中文摘要:
      目的 为探讨中枢神经系统辐射损伤的分子机理,采用寡聚核苷酸芯片技术研究16Gy60Coγ射线照射前后PC12细胞基因的表达变化。方法 采用大鼠寡聚核苷酸芯片技术进行辐射相关基因的筛查,芯片探针进行荧光交换标记,且每个基因重复3次点阵;分析结果,选取感兴趣基因进行RTPCR半定量验证。结果 基因芯片筛查有40个差异表达基因,其中上调基因37个,下调基因3个。这些差异基因主要涉及细胞周期、代谢、信号转导、免疫与应激、细胞增殖、生长发育及细胞凋亡等多个方面。RTPCR结果显示照射后48hPC12细胞OAS1、GARG16和IL6mRNA表达水平均上调,与芯片结果一致。结论 PC12细胞可用于神经细胞辐射后信号转导、细胞代谢、生长发育、应激反应以及细胞增殖等方面的研究。
英文摘要:
      Objective To explore the molecular mechanism of central nervous system(CNS) injury caused by ionizing radiation(IR) and to construct the gene expression profile database of IR-induced rat PC12 cells by microarray analysis. Methods Total RNAs of 16Gy IR-induced PC12 cells and normal PC12 cells were transcripted into cDNA by reverse transcription and labelled with cy5-dCTP and cy3-dCTP respectively.A total of 5705 oligo DNAs were selected for microarray analysis. Each sequence was dotted for three times repeatedly. RT-PCR was used to identify the interesting genes: OAS1, GARG16 and IL6. Results Of the 5705 genes tested, 40 genes showed changes in expression level with ratio higher than 2.0 and lower than 0.5.There were 37 genes showing up-regulation in radiated PC12 cells, whereas 3 genes showing down-regulation. Semi-quantitative RT-PCR confirmed that OAS1, GARG16 and IL6 mRNA expression levels were increased at 24 h and 48 h after 16 Gy 60Coγ-irradiation. Conclusion There are different gene expressions in the IR-induced PC12 cell model. The genes are involved in metabolism, cells cycle, immune response, signal transduction, and response to stress in PC12 cells after IR. These results provide deeper insight into the mechanism of CNS injury after IR.
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