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作 者:杨晖[1] 胡祖权[1] 董蓉[1] 薛慧[1] 龙金华[1] 李龙[1] 曾柱[1]
机构地区:[1]贵州医科大学生物与工程学院生物技术教研室,贵阳550004
出 处:《重庆医科大学学报》2015年第8期1071-1077,共7页Journal of Chongqing Medical University
基 金:国家自然科学基金资助项目(编号:81460254;11162003;31260227);贵州省留学回国人员科技活动基金资助项目(编号:2013-8);贵州省优秀青年科技人才支持计划资助项目(编号:2011-24);贵州省科学技术基金资助项目(编号:黔科合J字[2013]2058);贵州省科技计划资助项目(编号:黔科合LH字[2014]7092号);贵阳市科技计划项目资助项目(筑科合同[20141001]37号)
摘 要:目的:检测肝癌细胞微环境对人未成熟树突状细胞(immature dendritic cells,imDCs)基因表达谱的影响。方法:采用免疫磁珠阴性选择从人外周血分离CD14+单核细胞,在体外经粒-巨噬细胞采落刺激因子和白介素4诱导培养分化为im DCs,与Bel 7402肝癌细胞在Transwell中共培养48 h后,抽提imDCs的总RNA,利用人树突状细胞基因芯片检测其基因表达谱的变化。结果:与肝癌细胞共培养后,共有1 531个基因的表达发生改变(与对照组相比,变化表达倍数≥2),其中上调598个,下调933个,这些基因主要涉及信号转导、免疫应答、细胞黏附等功能。结论:肝癌细胞微环境能够影响imDCs多个基因的表达,这些基因的功能与imDCs的分化、成熟和功能有关,为进一步深入研究肝癌细胞对DCs免疫功能影响的分子机制奠定了基础。Objectives:To detect the influence of hepatocellular carcinoma cells micro-environment on the gene expression profiles in immature dendritic cells(im DCs). Methods:CD 14+ monocytes were collected by immunomagnetic beads from fresh harvested whole blood via negative selection. Im DCs were obtained by recombinant human granulocyte-macrophage colony-stimulating factor(rh GMCSF) and recombinant human inerleukins-4(rh IL-4) induction for 5 d in vitro. The collected im DCs were co-cultured with hepatoma cells Bel 7402. After 48 h,total RNA of the cells was collected,and human dendritic cells gene array was used to detect and analyze the changes of functional genes in im DCs. Results:After co-cultured with hepatoma cells,a total of 1 531 genes were changed(fold changes≥ 2 compared with that of control group),of which 598 up-regulated and 933 down-regulated. These genes were involved in signal transduction,immune response,cell adhesion,etc. Conclusion:The micro-environment of hepatocellular carcinoma can affect many genes regulation and expression of im DCs. These genes have influence on the differentiation,maturation and functions of imDCs. It is of significance to further understand the molecular mechanism of the hepatoma cells' influence on the immune function of imDCs.
分 类 号:R318.04[医药卫生—生物医学工程]
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