启动子甲基化调控NK-92MI细胞KIR3DL1基因表达  

Promoter methylation regulates KIR3DL1 gene expression in NK-92MI cell line

作　　者：高晓宁[1] 于力[1] 

机构地区：[1]解放军总医院血液科,北京100853

出　　处：《细胞与分子免疫学杂志》2008年第7期668-671,675,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30670898;30572108);国家重点基础研究发展计划(973)资助项目(2005CB522408)

摘　　要：目的:观察NK细胞系NK-92MI细胞中KIR3DL1基因启动子区的甲基化模式及去甲基化和组蛋白乙酰化对基因表达的影响,探讨KIR3DL1基因的表达调控机制。方法:采用亚硫酸氢盐测序法检测NK-92MI细胞中KIR3DL1基因启动子区的甲基化状况,应用甲基化抑制剂5-氮胞苷和(或)组蛋白去乙酰化转移酶抑制剂曲古抑菌素A处理NK-92MI细胞以诱导CpG岛去甲基化和组蛋白乙酰化,观察启动子区CpG岛甲基化和组蛋白乙酰化与KIR3DL1基因表达的关系。结果:NK-92MI细胞中KIR3DL1基因启动子区高甲基化,CpG二核苷酸甲基化频率在70%～100%之间;应用终浓度为2.5μmol/L和5μmol/L的5-氮胞苷作用72h可以诱导NK-92MI细胞中KIR3DL1mRNA表达量分别增加66.6%和114.6%;单用终浓度为50nmol/L的曲古抑菌素A不能诱导NK-92MI细胞中KIR3DL1mRNA表达量增加;此外,曲古抑菌素A和5-氮胞苷联用与单用5-氮胞苷相比也没有协同作用。结论:NK-92MI细胞中KIR3DL1基因表达受启动子甲基化调控。AIM： To investigate the methylation pattern of KIR3DLI promoter region in NK cell line NK-92MI and the effect of demethylation and histone acetylation on gene expression, and to study the possible regulation mechanism of KIR3 DLI expression. METHODS.. The methylation status of KIR3DLI promoter in NK-92MI cells was detected by bisulfite sequencing technique. Then NK-92MI cells were treated with the DNA-demethylating compound 5-azacytidine and （or） the inhibitor of histone deacetylase trichostatin A, and the expression of KIR3 DLI gene was observed. RESULTS： The CpG dinucleotides surrounding promoter region of KIR3DLI gene in NK-92MI cells were consistently methylated with a frequency of 70% - 100%. After 72 h treatment with 2.5 iJmol/L and 5 ijmol/L of 5-azacytidine, the mRNA expression of KIR3DLI gene in NK-92MI cells increased 66. 6% and 114.6%, respectively. However, after 72 h treatment with 50 nmol/L of trichostatin A, the mRNA expression of KIR3DLI gene in NK-92MI cells did not increase. Additionally, combined treatment with 5-azacytidine and tdchostatin A did not lead to a synergistic effect compared with 5- azacytidine alone. CONCLUSION： KIR3DLI expression in NK-92MI cells is regulated by promoter methylation.

关 键 词：杀伤细胞抑制性受体 基因表达调控 DNA甲基化 组蛋白乙酰化 

分 类 号：R392.11[医药卫生—免疫学]