硫化镍与苯并芘转化人支气管上皮细胞基因差异表达谱的研究

Study on differences of gene expression pattern in 16HBE cells transformationally treated with NiS and BPDE

目的 用cDNA微阵列技术探讨经硫化镍 (NiS)与二羟环氧苯并芘 (dihydroxyepoxybenzopyrene ,BPDE)转化后的人支气管上皮细胞 ( 16HBE)基因的差异表达 ;以了解不同种类和性质的化合物在转化细胞及致癌分子机制方面的异同 ;为预防人类生存环境不同外来化合物对人类健康的危害和影响提供科学依据。方法 取 16HBE分为NiS处理组 ,BPDE处理组和 16HBE(对照组 )三组同步培养 ,传代至第 3 5代 ;提取三种细胞的总RNA ,逆转录合成cDNA并以Cy3 dCTP和Cy5 dCTP荧光素分别标记制作探针。探针混合后与含 40 0 0个人类基因的芯片杂交。以ScanArray 40 0 0扫描仪扫描芯片 ,以GenPixPro 3 0软件分析荧光信号 ,对三组差异表达的基因进行生物学信息分析。结果 NiS转化的 16HBE细胞与BPDE转化的 16HBE细胞经分析比较 ,呈现差异表达的基因有 2 2个 ,其中 11个 ( 5 0 % )下调 ,另 11个 ( 5 0 % )上调。结论 NiS与BPDE对 16HBE细胞株的转化作用在抑癌基因 ,细胞周期蛋白相关基因 ,细胞凋亡和应激反应基因相关基因 ,DNA合成、修复和重组蛋白相关基因 ,DNA结合、转录和转录因子类基因 ,细胞受体相关基因 ,代谢相关基因 ,蛋白翻译和合成相关基因等多类基因上呈现差异表达。

Objective Using cDNA microarray to screen the differentially expressed genes in human bronchial epithelial cell line (16HBE) after transforming treatment with NiS and BPDE. Methods 16HBE cells were treated with NiS only (NiS group), treated with BPDE only (BPDE group) and untreated(control group). 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 4 000 human genes. The chips were scanned by ScanArray 4 000 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 22 genes exhibited differential expression between 16HBE cells treated only with NiS and BPDE. The expression of 11 genes (50%) was down-regulated and the other 11 (50%) was up-regulated. Conclusion The regulation of genes including tumor suppressor gene,apoptosis and stress response genes, immune related genes, DNA synthesis and repair genes, metabolism genes may be involved in transforming activity of NiS and BPDE. cDNA microarray technique is effective in screening the differentially expressed genes among different kinds of xenobiotics. Further analysis of those obtained genes will be helpful to understand the carcinogenic mechanism of xenobiotics in the levels of cell and molecular.