Central Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Reproductive Medical Center, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Nucleic Acid Ther. 2020 Dec;30(6):402-413. doi: 10.1089/nat.2020.0863. Epub 2020 Sep 29.
Colorectal cancer (CRC) is a prevalent malignancy with poor prognosis and survival. As a Ca binding protein, S100P plays a role in calcium-dependent signal transduction pathways that involve in diverse biological processes. Our previous studies have shown that S100P is overexpressed in CRC tissues and regulates cell growth, invasion, and metastasis in CRC. Therefore, S100P is expected to be an effective target for CRC therapy. Aptamers are short single-stranded oligonucleotides that could serve as specific and high-affinity probes to a wide range of target molecules for therapeutic purposes. In this study, we generated a novel DNA aptamer against S100P (AptS100P-1) by way of the SELEX process and high-throughput sequencing. The binding assay showed that AptS100P-1 had a high affinity for S100P protein. Further experiments indicated that AptS100P-1 is relatively stable in a cell culture system and could be used in flow cytometry analysis, dot blot assay, and fluorescence microscopy analysis to detect S100P. Moreover, AptS100P-1 was capable of binding to cells and had an inhibitory effect on CRC cell growth and Also, AptS100P-1 inhibited the migration and epithelial-mesenchymal transition of CRC cells expressing S100P. These results indicate a novel DNA aptamer targeting S100P, which might be a potential therapeutic strategy for targeting S100P against S100P-expressing CRC.
结直肠癌(CRC)是一种预后不良和生存率低的常见恶性肿瘤。作为一种 Ca 结合蛋白,S100P 在涉及多种生物过程的钙依赖性信号转导途径中发挥作用。我们之前的研究表明,S100P 在 CRC 组织中过表达,并调节 CRC 中的细胞生长、侵袭和转移。因此,S100P 有望成为 CRC 治疗的有效靶点。适体是短的单链寡核苷酸,可以作为针对各种治疗目的靶分子的特异性和高亲和力探针。在这项研究中,我们通过 SELEX 过程和高通量测序生成了针对 S100P 的新型 DNA 适体(AptS100P-1)。结合实验表明,AptS100P-1 对 S100P 蛋白具有高亲和力。进一步的实验表明,AptS100P-1 在细胞培养系统中相对稳定,可以用于流式细胞术分析、点印迹分析和荧光显微镜分析来检测 S100P。此外,AptS100P-1 能够与细胞结合,并对表达 S100P 的 CRC 细胞的生长具有抑制作用。同时,AptS100P-1 抑制表达 S100P 的 CRC 细胞的迁移和上皮-间充质转化。这些结果表明了一种针对 S100P 的新型 DNA 适体,它可能是针对 S100P 表达的 CRC 的潜在治疗策略。