Department of Pathophysiology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan 450001, China.
Henan Provincial Cooperative Innovation Center for Cancer Chemoprevention, Zhengzhou, Henan 450000, China.
Anal Chem. 2022 Dec 13;94(49):17212-17222. doi: 10.1021/acs.analchem.2c03863. Epub 2022 Dec 2.
Esophageal cancer, especially esophageal squamous cell carcinoma (ESCC), poses a serious threat to human health. It is urgently needed to develop recognition tools and discover molecular targets for early diagnosis and targeted therapy of esophageal cancer. Here, we developed several DNA aptamers that can bind to ESCC KYSE410 cells with a nanomolar range of dissociation constants by using cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX). The selected A2 aptamer is found to strongly bind with multiple cancer cells, including several ESCC cell lines. Tissue imaging displayed that the A2 aptamer can specifically recognize clinical ESCC tissues but not the adjacent tissues. Moreover, we identified integrin β1 as the binding target of A2 through pull-down and RNA interference assays. Meanwhile, molecular docking and mutation assays suggested that A2 probably binds to integrin β1 through the nucleotides of DA16-DG21, and competitive binding and structural alignment assays indicated that A2 shares the overlapped binding sites with laminin and arginine-glycine-aspartate ligands. Furthermore, we engineered A2-induced targeted therapy for ESCC. By constructing A2-tethered DNA nanoassemblies carrying multiple doxorubicin (Dox) molecules as antitumor agents, inhibition of tumor cell growth in vitro and in vivo was achieved. This work provides a useful targeting tool and a potential molecular target for cancer diagnosis and targeted therapy and is helpful for understanding the integrin mechanism and developing integrin inhibitors.
食管癌，尤其是食管鳞状细胞癌（ESCC），对人类健康构成严重威胁。迫切需要开发识别工具和发现分子靶点，用于食管癌的早期诊断和靶向治疗。在这里，我们通过基于细胞的配体指数富集系统进化（cell-SELEX）开发了几种 DNA 适体，它们可以与 ESCC KYSE410 细胞以纳摩尔范围的解离常数结合。所选择的 A2 适体被发现与多种癌细胞（包括几种 ESCC 细胞系）强烈结合。组织成像显示 A2 适体能特异性识别临床 ESCC 组织，但不能识别相邻组织。此外，我们通过下拉和 RNA 干扰实验鉴定整合素 β1 是 A2 的结合靶标。同时，分子对接和突变实验表明，A2 可能通过 DA16-DG21 的核苷酸与整合素 β1 结合，而竞争性结合和结构比对实验表明 A2 与层粘连蛋白和精氨酸-甘氨酸-天冬氨酸配体共享重叠的结合位点。此外，我们构建了 A2 诱导的 ESCC 靶向治疗。通过构建携带多个阿霉素（Dox）分子作为抗肿瘤剂的 A2 偶联 DNA 纳米组装体，实现了体外和体内肿瘤细胞生长的抑制。这项工作为癌症诊断和靶向治疗提供了一种有用的靶向工具和潜在的分子靶点，有助于理解整合素机制和开发整合素抑制剂。
