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

作　　者：高荣莲[1] 陈肖华[1] 饶亚岚[1] 张军权[1] 王治东[1] 董波[1] 侯丽莉[1] 张浩[1] 毛秉智[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》2005年第3期242-245,共4页Chinese Journal of Radiological Medicine and Protection

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。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 ()^(60)Co γ-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.;

关 键 词：PC12细胞 差异表达基因 寡聚核苷酸 芯片技术 电离辐射 大鼠 中枢神经系统辐射损伤 ^60COΓ射线照射 RT-PCR 筛选 mRNA表达 信号转导 细胞增殖 生长发育 分子机理 表达变化 细胞基因 相关基因 芯片探针 基因重复 分析结果 

分 类 号：R818[医药卫生—放射医学]