【作者】 宾晓农； 谭敏； 吕嘉春； 蒋义国； 陈家堃；

【Author】 Bin Xiaonong Lu Jiachun Chen Jiakung Ji Weidong Tan Min.Guangzhou medical college, Guangzhou,510182,China

【机构】 广州医学院化学致癌研究所； 广东省中医院；

【摘要】 目的:应用基因芯片技术筛选二羟环氧苯并芘(BPDE)转化细胞的相关基因,探讨基因芯片技术高通量筛查在细胞、分子毒理学研究中的应用价值。方法:按一步法抽提实验组BPDE转化支气管上皮细胞和对照组正常支气管上皮细胞的总RNA并纯化mRNA;将4096种人类基因PCR产物用Cartesia Pixsys8000点样仪按微矩阵排列点样于化学涂层的载玻片上,制成基因芯片;将等量的BPDE转化支气管上皮细胞和正常支气管上皮细胞的mRNA分别逆转录合成荧光分子掺入的cDNA一链做探针,混合后杂交上述基因芯片。经严格洗片后用ScanArray4000扫描仪扫描芯片荧光信号图象,计算机分析后比较BPDE转化支气管上皮细胞和正常支气管上皮细胞中差异表达的基因。结果:在4096种人类基因中,BPDE转化支气管上皮细胞和正常支气管上皮细胞间存在差异表达的基因有143条,其中高表达52条,低表达91条。结论:基因芯片技术在筛选BPDE转化细胞相关基因的改变上,具有高通量、高敏感、快速等特点,对毒物在细胞、分子毒作用和致癌机理的研究中意义重大。更多还原

【Abstract】 Objective Using cDNA microarray to screen for the differentially expressed genes in human bronchial epithelial cell line 16HBE after treatment with BPDE.Methods 16HBE cells and 16HBE cells were treated with BPDE only.The total RNAs were extracted from these cells.The cDNA probes were prepared by labeling with Cy3-dCTP and Cy5-dCTP respectively through reverse transcription.The mixed probes were then hybridized to the cDNA microarray chips containing 4096 human genes.The chips were scanned by ScanArray 4000 laser scanner.The acquired fluorescent signals were analyzed by GenPix Pro 3.0 software.Bioinformatical analysis of those differentially expressed genes had been performed.Results 143 genes exhibited differential expression between cells treated with BPDE only and 16HBE.The expression of 91 genes(63.63%) was down-regulated and of 52 genes(36.36%) was up-regulated.Conclusion The regulation of genes including stress response genes,immune related genes,DNA synthesis and repair genes, metabolism genes may be involved in itransforming activity of NiS.更多还原