去甲基化处理对NK-92MI细胞系抑制性受体KIR表达的影响  

Effect of Demethylation Treatment on the Expression of Inhibitory Receptor KIR Gene in NK-92MI Cell Line

作　　者：高晓宁[1] 林季[2] 王莉莉[1] 高丽[1] 靳海杰[1] 孙敬芬[1] 于力[1] 

机构地区：[1]解放军总医院血液科,北京100853 [2]解放军总医院基础医学研究所生化室,北京100853

出　　处：《中国实验血液学杂志》2009年第3期656-660,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(编号30670898;30572108);国家重点基础研究发展计划(973)资助项目(编号2005CB522400)

摘　　要：为分析NK细胞系NK-92MI细胞中抑制性杀伤细胞免疫球蛋白样受体(killer cell immnoglobulin-like re-ceptor,KIR)的基因启动子区的甲基化模式及去甲基化处理对基因表达的影响,探讨基因kir可能的表达调控机制,采用亚硫酸氢盐测序法检测NK-92MI细胞kir2DL1和kir2DL2/2DL3基因启动子区的甲基化水平,应用甲基化抑制剂5-氮胞苷处理NK-92MI细胞以诱导CpG岛去甲基化,RT-PCR方法检测其基因表达。结果显示:kir2DL1和kir2DL2/2DL3基因启动子区CpG二核苷酸甲基化频率依次在25%-88%、5%-80%之间;应用甲基化抑制剂5-氮胞苷可以诱导NK-92MI细胞重新表达kir2DL1基因,并使kir2DL1、kir2DL2和kir2DL3基因表达量增加。结论:启动子甲基化参与调控NK-92MI细胞基因kir表达。The aim of this study was to analyze the promoter methylation patterns of inhibitory killer cell immunoglobulin-like receptor （KIR） which gene expression and the effect of demethylation treatmant were studied, and to explore the possible regulation mechanism of inhibitory kir gene expression. The promoter methylation levels of kir2DL1 and kir2DL2/kir2DL3 in NK-92MI cell line were detected by bisulfite sequencing technique. Then NK-92MI cells were treated with 5-azacytidine to induce the demethylation of CpG islands. The levels of gene expression of kir were determined by RT-PCR. The results demonstrated that the methylation frequencies of CpG dinucleotides surrounding the promoter regions of kir2DL1 and kir2DL2/kir2DL3 genes were 25% to 88% and 5% to 80% respectively. DNA-demethylating treatment with 5-azacytidine resulted in re-expression of kir2DLl gene and increased expressions of kir2DL1, kir2DL2 and kir2DL3 genes in NK-92MI cells. In conclusion, the promoter DNA methylation participates in the regulation of kir gene expression in NK-92MI cells.

关 键 词：杀伤细胞免疫球蛋白样受体 NK-92MI细胞系 去甲基化处理 

分 类 号：R392.11[医药卫生—免疫学]