电离辐射相关低密度寡核苷酸基因芯片制备.docVIP

电离辐射相关低密度寡核苷酸基因芯片制备 　 作者：郭万峰，王升启，黄坚，郭国祯 【关键词】 肺癌 Fabrication of lowdensity ionizing radiationrelated oligonucleotide microarray 　　【Abstract】 AIM: To fabricate an oligonucleotide microarray applied to ionizing radiation. METHODS: According to the genes reported previously in radiation responses and their family genes or related genes, oligonucleotide microarray was designed. mRNA sequences of these genes were obtained from the GenBank. Specific probes were designed with Mprobe software and synthesized by AB18909 DNA synthesis system and spotted onto aldehydecoated glass slides using Cartesian Pix sys 3000 spotter in our laboratory. Using this microarray, we identified genes showing altered expression in different radiosensitive lung cancer cell lines. To provide independent confirmation of microarray data, semiquantitative RTPCR was performed on four genes. RESULTS: Lowdensity radiationrelated oligonucleotide microarray was fabricated successfully, which had 143 genes including 127 involved in stress response, cell cycle progression, DNA damage and repair, apoptosis and proliferation, together with 11 housekeeping genes and 3 luciferase genes as positive controls and 2 rice genes as negative controls. Eighteen differentially expressed genes were screened by this microarray. Four genes were detected by RTPCR, and the results were in accordance with those from the microarray data, even though the value for each mRNA level might be different between the two measurements. CONCLUSION: The radiationrelated oligonucleotide microarray can be used to investigate the differentially expressed genes in different radiosensitive cancer cells and provide a platform for radiation research. 　　【Keywords】 microarray; ionizing radiation; lung neoplasms; cell line, tumor 　　【摘要】 目的: 构建一种适用于辐射研究领域的低密度寡核苷酸基因芯片. 方法: 根据以往的研究报告，筛选参与辐射生物效应的基因和相关基因，从GenBank获取mRNA序列，使用Mprobe软件设计特异的探针，构建低密度辐射相关的基因芯片.在不同辐射敏感性肺癌细胞系中对该芯片的灵敏度和特异性进行评价，并对一些差异表达基因进行RTPCR验证，以确认芯片检测结果的可信性. 结果: 构建出了辐射相关的寡核苷酸基因芯片,该芯片共含有143