microRNA调控NK细胞KIR3DL1表达初探

microRNA regulates KIR3DL1 expression in natural killer cells

目的初步探寻人外周血自然杀伤细胞(Nature Killer,NK)杀伤细胞免疫球蛋白样受体(Killer Cell Immnoglobulin-Like Receptor,KIR3DL1)表达可能存在的microRNA(miR)调控机制。方法利用生物信息学方法,从miR信息库中筛选出可能与KIR3DL1相关的miRs,构建含KIR3DL1 3’非翻译区(UTR)的PGL3质粒,分别将含相应miR的PcDNA3.0质粒与前者共转染293T细胞,通过荧光素酶报告实验及之后的突变实验筛选出可能调控KIR3DL1的miR。结果通过TARGET SCAN信息库筛选了miR-146b等10个miR;转染miR-146b后,荧光素酶活性下降最多(61.3%),突变其KIR3DL1 3’UTR靶位点后荧光素酶活性恢复(91.4%)。结论 miR-146b可与KIR3DL1’UTR在靶位点特异结合,很可能参与KIR3DL1表达的调控。

Objective To study the potential mechanism of microRNA（miR）underlying the regulation of KIR3DL1 expression in human peripheral blood derived natural killer（NK） cells.Methods KIR3DL1-associated miR was searched from miR database using bioinformatics techniques.PGL3 plasmids containing KIR3DL13＇UTR were constructed.PcDNA3.0 containing corresponding miR and KIR3DL13＇UTR were co-transfected into 293T cells.miR that may regulate KIR3DL1 expression before and after mutation was detected by luciferase reporter assay.Results Ten miRs including miR-146b were searched from the Target Scan Database.After transfection of miR-146b,the activity of luciferase was decreased to 61.3%.After mutation of the KIR3DL1 3＇UT,the activity of luciferase increased 91.4%.Conclusion miRNA 146b can specifically bind to KIR3DL13＇UTR and participate in regulation of KIR3DL1 expression.