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基于细胞的单链 DNA 适体特异性靶向胃癌。

A cell-based single-stranded DNA aptamer specifically targets gastric cancer.

出版信息

Int J Biochem Cell Biol. 2014 Jan;46:1-8.

Abstract

Gastric cancer is one of the most prevailing cancers with high morbidity and mortality. Limitations in the current diagnosis and therapy, specially lacking of specific molecular therapeutic targets, ask for the development of new strategies. Aptamer, a newly developed adaptive molecule, could be used in clinical detection and therapy because of its high affinity and specificity. As no aptamer has ever been developed in preventing gastric cancer so far, we were the first who cloned such an aptamer specifically targeting gastric cancer. The aptamer was selected by systematic evolution of ligands by exponential enrichment with gastric cancer cell-line HGC-27 as target cell line and immortalized gastric epithelial cell-line GES-1 as control cell line. The affinity and specificity of candidate aptamers were examined by flow cytometry, confocal imagining and aptamer-based histochemistry staining. After 19 cycles of systematic evolution of ligands by exponential enrichment and subsequent cloning and sequencing, an aptamer with the highest affinity and specificity (nominated as AGC03) among candidates was screened out from a random single-stranded DNA pool. Moreover, AGC03 could not only specifically bind to gastric cancer cells (the equilibrium dissociation constant value was 16.49 ± 0.40 nM) in vitro, but also recognize cancer cells in human cancer tissue. Our most important finding is that AGC03 could even be internalized into cells automatically. In conclusion, we obtained a novel aptamer specifically targeting gastric cancer,which is an effective tool for both gastric cancer diagnosis and drug delivery.

摘要

胃癌是最常见的癌症之一,具有高发病率和死亡率。目前的诊断和治疗方法存在局限性,特别是缺乏特异性的分子治疗靶点,因此需要开发新的策略。适体是一种新开发的适应性分子,由于其高亲和力和特异性,可用于临床检测和治疗。由于迄今为止还没有开发出用于预防胃癌的适体,我们是第一个专门针对胃癌克隆这种适体的人。该适体通过指数富集的配体系统进化筛选,以胃癌细胞系 HGC-27 为靶细胞系,永生化胃上皮细胞系 GES-1 为对照细胞系进行筛选。通过流式细胞术、共聚焦成像和基于适体的组织化学染色来检查候选适体的亲和力和特异性。经过 19 轮配体系统进化指数富集和随后的克隆和测序,从随机单链 DNA 库中筛选出一种亲和力和特异性最高的适体(命名为 AGC03)。此外,AGC03 不仅可以在体外特异性结合胃癌细胞(平衡解离常数值为 16.49±0.40 nM),还可以识别人类癌组织中的癌细胞。我们最重要的发现是,AGC03 甚至可以自动内化进入细胞。总之,我们获得了一种新型的、专门针对胃癌的适体,它是胃癌诊断和药物输送的有效工具。

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