基因芯片对ibp缺失后激活t淋巴细胞基因表达-第三军医大学学报.doc

IBP表达抑制后活化T淋巴细胞基因表达谱变化分析 向 莉，李 鹏，张竹君，杨明珍，陈 安，胡川闽 （400038重庆，第三军医大学医学检验系临床生化教研室）［摘要］ 目的 利用全基因组寡核苷酸芯片检测干扰素调节因子4结合蛋白（IRF-4 binding protein，IBP）后T淋巴细胞基因表达谱的差异。方法 采用本室构建的IBP表达抑制的Jurkat细胞及相应对照细胞为对象，Anti-CD3、CD28 mAb处理细胞后提取刺激2448 h的细胞RNA，利用北京博奥生物有限公司22K Human Genome Array芯片检测基因表达的差异以MAS.doc.V1软件，结合KEGG、NCBI等生物信息学数据库检索分析差异表达基因功能和网络关系。结果IBP基因表达抑制的Jurkat细胞在TCR信号刺激2448 h后，细胞能量代谢、周期生长、转录调控及凋亡等多种类型的基因发生差异表达改变，时相组共有56个差异表达趋势一致基因，其中个基因共同上调，个基因共同下调。结论在Jurkat T细胞IBP表达抑制所致多个基因差异表达。［关键词］干扰素调节因子4结合蛋白；T淋巴细胞；基因芯片［中图分类号］R392. 11 ［文献标码］A Gene expression profile of activated T lymphocyte with IBP deficient by oligonucleotide microarray Xiang Li, Li Peng, Zhang zhujun, Yang mingzhen, Chen an, Hu Chuanmin （Department of Clinical Biochemistry, College of Medical LaboratoryThird Military Medical University, Chongqing , 400038, China） [Abstract] Objective　To investigate the difference of gene expression between activated IBP-deficient and wild T lymphocyte by oligonucleotide microarray and provide basis for further studying the role of IBP in T lymphocyte activation. Methods　Total RNA was isolated from IBP-deficient and its parental jurkat T cells with anti-CD3 and anti-CD28 mAb stimulation, and synthesized into double-stranded cDNA that was then synthesized into biotin-labeled cRNA probe by in vitro transcription. The cRNA probes were separately hybridized with 22K Human Genome Array Chip, and the signals were scanned by the GeneArray Scanner. The results were analyzed by bioinformatics. Results　In comparison with the gene expression profile of parental Jurkat T cells, 17 genes were up-regulated and 39 genes down-regulated in activated IBP-deficient Jurkat T cells. According to Gene Ontology and Tree View analysis, these genes were involved in energy metabolism, cell cycle, transcription and apoptosis and so on. Conclusion　Many differential expression genes induced by the lost of IBP in Jurkat T cells have been screened by high-throughput gene chip method. Validating their mo