Fabrication of lowª²density ionizing radiationª²related oligonucleotide microarray

    GUO Wanª²Feng, WANG Shengª²Qi, HUANG Jian, GUO Guoª²Zhen

    1Department of Radiation Medicine, School of Preventive Medicine, Fourth Military Medical University, Xi¬ðan 710032, China, 2PLA Key Laboratory of Biochips, Institute of Radiation Medical Science, Academy of Military Medical Sciences, Beijing 100850, China

    ¡¾Abstract¡¿ 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 aldehydeª²coated 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, semiª²quantitative RTª²PCR was performed on four genes. RESULTS: Lowª²density radiationª²related 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 RTª²PCR, 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 radiationª²related oligonucleotide microarray can be used to investigate the differentially expressed genes in different radiosensitive cancer cells and provide a platform for radiation research. ......