显性调控相关早反应基因的大规模扫描和分析

A large-scale scanning and comparative analysis of early response genes associated with dominant regulation

目的 动态观察白细胞介素 1 0 (IL 1 0 )作用后 ,淋巴细胞早反应基因表达谱变化 ,为下一步评价患者免疫状态和诱导治疗提供理论依据。方法 应用DNA微矩阵技术 ,分离正常人外周血单个核细胞 (PBMC) ,予以抗CD3McAb及CD3McAb +IL 1 0刺激 ,抽提 2 4hPBMCmRNA ,反转录后与芯片杂交 ,通过生物信息学方法比较分析。结果 培养 2 4h后 ,抗CD3McAb +IL 1 0组与抗CD3McAb组比较有 1 5个基因发生改变 ,5个基因表达明显下调 ,1 0个基因上调 ,主要功能在于促进基因编码区连接 ,影响细胞骨架和运动 ,不同部位的磷酸化、去磷酸化 ,细胞识别 ,会促进细胞生长作用 ,细胞凋亡 ,信号传递的基因则重点涉及钙离子 -钙调蛋白、G 蛋白信号通路。结论 在IL 1 0作用下 ,一系列基因转录活化 ,其中许多先前与免疫细胞无明确功能的相关基因参与了T细胞活化与调控 ,是否该群基因即代表IL 1 0作用下淋巴细胞的基因表达标签 ,尚须进一步证实 ;要完全明确T细胞调控网络 ,还应全面分析T淋巴细胞基因表达谱 ,从中筛选差异有显著意义的基因 ,进行功能研究。

Objective To scan and comparatively analyze the genes associated with dominant regulation after the action of interleukin 10 (IL 10) by microarray, especially early response genes.Methods The peripheral blood mononuclear cells (PBMC) were isolated, stimulated with anti cd3 monoclonal antibody or both anti cd3 monoclonal antibody and IL 10. The mRNA of PBMC at 24?h were isolated and purified, reversely transcripted and hybridized with gene chips. The results were analyzed by bioinformatics.Results After culturing for 24?h, 15 genes changed in the anti CD3McAb+IL 10 group as compared with those in the anti CD3McAb+IL 10 group: the expression of 5 genes was obviously downregulated and 10 upregulated. Among them, some were related with cell skeleton and motor protein, signal transduction, cell recognition, promoting cell growth and metabolism etc, and others were undefined genes.Conclusions After the gene expression profile of lymphocyte scanned under the action of IL 10 by microarray, many genes that were not previously connected to immune regulation were revealed. Further identification awaits to verify whether the genes represent expression tag of the action of IL 10. The results implicated that it is essential to completely analyze the genes profile for understanding the function and regulation of lymphocytes.