基于细胞的 SELEX 技术筛选和鉴定识别人肝癌的 DNA 适体。
Cell-SELEX based selection and characterization of DNA aptamer recognizing human hepatocarcinoma.
机构信息
Institute of Nature and Environmental Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
出版信息
Bioorg Med Chem Lett. 2013 Mar 15;23(6):1797-802. doi: 10.1016/j.bmcl.2013.01.040. Epub 2013 Jan 22.
Single-stranded DNA aptamers recognizing human hepatocarcinoma were isolated by means of a systematic evolution of ligands by exponential enrichment using whole cells as targets (cell-SELEX). After 11 rounds of cell-SELEX procedure using human hepatoma HepG2 cells as targets and human normal hepatocyte cells as counterparts, 12 independent DNA aptamer candidate sequences were obtained. The specific interaction between selected DNA aptamers and targeted cell was confirmed. Dissociation constants of the 12 sequences obtained were also estimated in the range of 19-450nM. Moreover, the consensus secondary structure was found in the isolated aptamers, which was responsible to the recognition of HepG2 cells.
通过以全细胞为靶标的指数富集配体系统进化技术（cell-SELEX），分离出与人肝癌有特异性识别的单链 DNA 适体。经过 11 轮以人肝癌 HepG2 细胞为靶标、人正常肝细胞为对照的 cell-SELEX 筛选，得到了 12 个独立的 DNA 适体候选序列。鉴定了筛选出的 DNA 适体与靶细胞之间的特异性相互作用，并对获得的 12 个序列的解离常数进行了估计，范围在 19-450nM 之间。此外，还发现分离出的适体具有保守的二级结构，这是其识别 HepG2 细胞的基础。
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