CpG-寡核苷酸对免疫细胞基因表达的影响

目的:研究CpG-寡核苷酸(CpG-ODN)对免疫细胞基因表达的影响.方法:用Cy3和Cy5两种荧光分子通过逆转录反应将CpG-ODN刺激前后的单核-巨噬细胞系RAW264.7细胞的mRNA分别标记成探针,混合后与小鼠基因表达谱芯片杂交,通过扫描、计算机分析,寻找经CpG-ODN刺激后差异表达的基因.结果:CpG-ODN刺激RAW264.7细胞6 h后,2次芯片杂交筛选出重复差异表达的基因119条,其中74条上调,45条下调;这些基因涉及细胞周期、免疫调控、脂肪代谢、泡沫细胞形成、信号转导等过程.结论:CpG-ODN可广泛调控巨噬细胞内多种基因的表达,对相关基因群的进一步研究有助于深入了解CpG-ODN的作用机制.

Effects of CpG oligodeoxynucleotide on gene expression in immunocytes

Objective: To study the influence of CpG oligodeoxynucleotide (CpG-ODN) on the gene expression in im-munocytes and its mechanisms. Methods: RAW264. 7 cells were stimulated with CpG-ODN for 6 h,and mRNA from both control and treated cells were isolated and purified,then were reversely transcribed to cDNA with the incorporation of fluorescent-labeled dUTP to prepare the hybridization probes. The mixed probes were then hybridized to the cDNA microarray MGEC-80s. After high-stringent washing, the cDNA microarray was scanned by computer system and the differently expressed genes were obtained. Results: A total of 119 differently expressed genes were detected after CpG-ODN stimulation,of which 74 were up-regulated and 45 were down-regulated. These genes were related to cell cycle,immune modulation,lipid metabolism,foam cell formation,and signal transduction. Conclusion: CpG-ODN may widely modulate the expression of many genes in immune cells,and further analysis of related genes may help understand the molecular mechanisms of CpG-ODN.