小鼠microRNA-135a与STAT6靶点作用的体外研究  

Target Effect of Murine MicroRNA-135a and STAT6 in Rats in Vitro

作　　者：罗燕云[1] 阳光[1] 张钊[1] 陈睿尧 陶泽璋[2] 

机构地区：[1]广州军区武汉总医院耳鼻喉科,湖北武汉430070 [2]武汉大学人民医院耳鼻咽喉-头颈外科

出　　处：《华南国防医学杂志》2016年第1期1-4,13,共5页Military Medical Journal of South China

基　　金：国家自然科学基金项目(81070766)

摘　　要：目的结合TargetScan软件预测,体外验证小鼠microRNA-135a(miR-135a)与信号传导及转录激活因子6(signal transducer and activator of transcription 6,STAT6)的靶点结合作用。方法将聚合酶链反应(polymerase chain reaction,PCR)扩增的靶基因STAT6的非编码3'UTR和突变基因分别克隆到双荧光素酶报告载体中,构建野生型和突变型载体。分别将miR-135a模拟物、模拟对照物与靶基因野生型、突变型质粒载体共转染体外培养的293T细胞,测定载体的校正荧光hluc值及报告荧光hRluc值。结果与模拟对照物相比较,miR-135a模拟物与靶基因STAT6的3'UTR野生型质粒的相互作用能明显降低hRluc/hluc的相对荧光比值,差异有统计学意义(P<0.01);而miR-135a模拟物与突变型质粒相作用则对hRluc/hluc的相对荧光比值比较差异无统计学意义(P>0.05)。结论小鼠miR-135a与STAT6的3'UTR体外特异性结合作用,为进一步探讨小鼠miR-135a对变应性鼻炎的免疫调控机制提供了依据。Objective To validate the target effect of murine microRNA-135a（miR-135a）and signal transducer and activator of transcription 6（STAT6）in vitro combined the TargetScan software.Methods The amplified STAT63＇UTR target gene and mutant gene by polymerase chain reaction（PCR）were cloned respectively into dual-luciferase reporter vector to build the wild and mutant type carriers.The 293 T cells cultured in vitro were co-transfected respectively by miR-135 a mimic and mimic control with the wild and mutant carriers.Corrected fluorescence hluc value and report fluorescence hRluc value were measured.Results Compared with mimic control the mutual response between miR-135 a mimic and the wild carrier with STAT6 3＇UTR target gene could significantly decrease the relative hRluc/hluc fluorescence ratio（P〈0.01）,while the mutual response between miR-135 a mimic and the mutant carrier didn＇t（P〈0.05）.Conclusion The specific binding effect of murine miR-135 a and STAT6 3＇UTRin vitro provides a certain proof for the further exploration of the immune regulation mechanism of murine miR-135 a in allergic rhinitis.

关 键 词：MICRORNA 信号传导及转录激活因子 模拟物 小鼠 变应性鼻炎 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]