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Screening and characterization of an Annexin A2 binding aptamer that inhibits the proliferation of myeloma cells
Biochimie ( IF 3.3 ) Pub Date : 2018-06-15 , DOI: 10.1016/j.biochi.2018.06.003
Weihua Zhou , Yibin Zhang , Yayue Zeng , Minyuan Peng , Hui Li , Shuming Sun , Bianying Ma , Yanpeng Wang , Mao Ye , Jing Liu

Multiple myeloma (MM) is a malignant plasma cell disease and is considered incurable. Annexin A2 (ANXA2) is closely related to the proliferation and adhesion of MM. Using protein-SELEX, we performed a screen for aptamers that bind GST-ANXA2 from a library, and GST protein was used for negative selection. The enrichment of the ssDNA pool was monitored by filter-binding assay during selection. After nine rounds of screening and high-throughput sequencing, we obtained six candidate aptamers that bind to the ANXA2 protein. The affinities of the candidate aptamers for ANXA2 were determined by ELONA. Binding of aptamer wh6 to the ANXA2 protein and to the MM cell was verified by aptamer pulldown experiment and flow cytometry, respectively. Aptamer wh6 binds the ANXA2 protein with good stability and has a dissociation constant in the nanomolar range. The binding specificity of aptamer wh6 was confirmed in vivo in nude mouse xenografts with MM cells and with MM bone marrow aspirates. Furthermore, aptamer wh6 can block MM cell adhesion to ANXA2 and block the proliferation of MM cells induced by ANXA2. In summary, wh6 can be considered a promising candidate tool for MM diagnosis and treatment.



中文翻译:

抑制骨髓瘤细胞增殖的膜联蛋白A2结合适体的筛选和表征

多发性骨髓瘤(MM)是一种恶性浆细胞疾病,被认为是无法治愈的。Annexin A2(ANXA2)与MM的增殖和粘附密切相关。使用蛋白质SELEX,我们从文库中筛选了与GST-ANXA2结合的适体,GST蛋白用于阴性选择。在选择过程中通过滤膜结合测定法监测ssDNA库的富集。经过九轮筛选和高通量测序,我们获得了六种与ANXA2蛋白结合的候选适体。通过ELONA确定ANXA2的候选适体的亲和力。分别通过适体下拉实验和流式细胞术验证了适体wh6与ANXA2蛋白和MM细胞的结合。适体wh6以良好的稳定性结合ANXA2蛋白,并且在纳摩尔范围内具有解离常数。在具有MM细胞和MM骨髓抽吸物的裸鼠异种移植物中,体内证实了适体wh6的结合特异性。此外,适体wh6可阻断MM细胞对ANXA2的粘附并阻断由ANXA2诱导的MM细胞的增殖。总而言之,wh6可以被认为是用于MM诊断和治疗的有前途的候选工具。

更新日期:2018-06-15
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