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去甲基化处理对NK-92MI细胞系抑制性受体KIR表达的影响
引用本文:高晓宁,林季,王莉莉,高丽,靳海杰,孙敬芬,于力. 去甲基化处理对NK-92MI细胞系抑制性受体KIR表达的影响[J]. 中国实验血液学杂志, 2009, 17(3): 656-660
作者姓名:高晓宁  林季  王莉莉  高丽  靳海杰  孙敬芬  于力
作者单位:解放军总医院血液科,北京,100853
基金项目:国家自然科学基金面上项目,国家重点基础研究发展规划(973计划) 
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(killercellimmnoglobulin-likere—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基因启动予区的甲基化水平,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。

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

Effect of Demethylation Treatment on the Expression of Inhibitory Receptor KIR Gene in NK-92MI Cell Line
GAO Xiao-Ning,LIN Ji,WANG Li-Li,GAO Li,JIN Hai-Jie,SUN Jing-Fen,YU Li. Effect of Demethylation Treatment on the Expression of Inhibitory Receptor KIR Gene in NK-92MI Cell Line[J]. Journal of experimental hematology, 2009, 17(3): 656-660
Authors:GAO Xiao-Ning  LIN Ji  WANG Li-Li  GAO Li  JIN Hai-Jie  SUN Jing-Fen  YU Li
Affiliation:(Department of Hematology, PLA General Hospital, Beijing 100853, China; 1Laboratory of Biochemistry, Institute of Basic Medical Sci- ences, PLA General Hospital, Beijing 100853, China)
Abstract:
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.
Keywords:killer cell immnoglobulin-like receptor  NK-92MI cell line  demethylation treatment
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