新型HER2适配体的筛选及鉴定  

Identification of a novel DNA aptamer against HER2

作　　者：刘喆[1] 杨先达[1] 

机构地区：[1]中国医学科学院基础医学研究所,北京100005

出　　处：《基础医学与临床》2012年第6期608-612,共5页Basic and Clinical Medicine

基　　金：国家重大科学计划(2011CB933504;2011CB911003;2011CB911004);国家自然科学基金(81071870)

摘　　要：目的研发能特异性结合人表皮生长因子受体2(HER2)的DNA适配体,为开发针对HER2的新型肿瘤靶向诊疗技术提供依据。方法体外合成全长86个碱基并含有40个随机寡核苷酸的单链DNA文库,以HER2表位肽为靶标,利用指数富集的配体系统进化技术(SELEX),从单链DNA文库中筛选能够选择性结合HER2多肽的核酸适配体;流式细胞术检测富集进度、适配体与HER2蛋白及HER2阳性细胞的结合特性;MFold软件预测二级结构。结果经多轮筛选获得了能够识别HER2多肽的DNA适配体HA5,其能够选择性地结合HER2蛋白及HER2阳性的乳腺癌细胞,而不结合胰蛋白酶和HER2阴性细胞。结论 DNA适配体HA5能选择性地识别HER2阳性的乳腺癌细胞,在研发针对HER2的新型肿瘤靶向诊疗技术方面具有应用潜能。Objective To develop HER2 DNA aptamers that may potentially serve as a tumor-binding ligand in novel HER2-targeted therapeutic strategies.Methods A single-stranded 86nt DNA library containing 40 random oligonucleotides was synthesized in vitro.A new aptamer HA5 was developed by SELEX technique with HER2 epitope peptide as target.Flow cytometry was performed to monitor the enrichment of aptamer pool and to evaluate the binding characterization of HA5.The structure of HA5 was predicted by MFold.Results The selected HA5 bound to the HER2 and had minimal cross reactivity to trypsin.In addition,the aptamer was found to preferentially bind to HER2-positive but not to HER2-negative cells.Conclusions HA5 may have application potentials in targeted therapy against HER2-positive malignancies.

关 键 词：适配体 SELEX HER2 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]