脂多糖活化巨噬细胞的数字基因表达谱分析

Digital gene expression profile of RAW264. 7 macrophages stimulated with lipopolysaccharide

目的研究巨噬细胞经脂多糖(LPS)处理后基因表达谱的变化。方法 LPS 0.01 mg·L-1作用RAW264.7细胞24 h,应用数字基因表达谱(DGE)技术筛选差异表达基因,分析差异基因富集的Gene ontology(GO)功能和Kyoto encyclopoedia of genes and genomes(KEGG)信号通路,并通过定量PCR对部分差异表达基因进行验证。结果 LPS处理后,共有1959个基因差异表达,其中上调974个,下调985个。GO分析显示,差异表达基因主要发挥蛋白结合和催化活性等分子功能,以及调节细胞过程、对环境刺激的反应和免疫系统等生物学功能。信号通路分析显示,显著富集的途径与炎症、免疫系统和癌症等疾病密切相关。结论 DGE技术验证了LPS介导炎症反应的NF-κB、丝裂原激活蛋白激酶和Jak-STAT 3条主要的信号转导途径,发现了细胞周期、细胞凋亡和细胞外识别等新的途径。

OBJECTIVE To research on the systematic transcriptomic changes of RAW264.7 macrophages stimulated with lipopolysaccharides （LPS）. METHODS RAW264.7 cells were treated with LPS 0.01 mg·L-1 for 24 h, the transcriptome profiling of the differentially expressed genes was analyzed by digital gene expression tag profiling （DGE） technology. The genes were analyzed with Gene Oncology （GO） Term and Kyoto Encyclopedia of Genes and Genomes （KEGG） to screen the signals significantly enriched. The genes differently expressed were verified with the real-time quantitative PCR. RESULTS There were 1959 genes expressed significantly difference. 974 genes were up-regulated and 985 genes were down-regulated. GO analysis indicated that differentially expressed genes mainly exercised the molecular functions of catalytic activity and protein binding, and involved in biological process of biological regulation, response to stimulus and immune system process. Pathway enrichment analysis showed that these genes involving in immune response, inflammation and cancer were significantly enriched. CONCLUSION The DGE technology verifies that LPS can stimulate the production of cytokines by activating of three classic inflammatory signal pathways, including mitogen-activated protein kinase, NF-κB and Jak-STAT pathways. From DGE results, some new pathways, including cell cycle, cell apoptosis and cell recognization signal pathways etc are also revealed.