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靶向具有恶性潜能的结肠癌细胞的新型DNA适配体的筛选与鉴定

Selection and characterization of novel DNA aptamers targeting colorectal cancer cells with malignant potential.

作者信息

Ma Yinuo, Wang Qun, Sun Shihan, Li Yanxi, Wang Rui, Yang Haihui, Wang Dian, Hao Yangyang, Tong Yujing, Li Wanming

机构信息

Department of Cell Biology, Key Laboratory of Cell Biology, Ministry of Public Health, Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, 110122, PR China.

Department of Colorectal Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang, 110042, PR China.

出版信息

Anal Bioanal Chem. 2025 Sep 2. doi: 10.1007/s00216-025-06091-9.

Abstract

Colorectal cancer (CRC) is associated with high morbidity and mortality rates worldwide. Therefore, early diagnosis and treatment are of great significance for improving the survival rate of CRC patients. Herein, we performed subtractive cell-SELEX using two cell lines (LS174T and CL187) with the same genetic background but different malignant potentials for the target and negative cells. Two aptamers LS1 and LS52 showed high affinity with target LS174T cells and high specificity for CRC cells with malignant potential, and exhibited good biological stability. In addition, we found that aptamer LS1 had the effect of promoting in vitro proliferation and monoclonal formation of CRC cells, and upregulated the expression of proliferation related proteins. Based on the high affinity of aptamer LS52, using LS52 as a molecular probe could effectively distinguish clinical CRC tissues and adjacent normal tissues, CRC metastatic tissues and non-metastatic tissues. Analysis of protein expression in magnetic bead sorted cells showed that the expression of malignant potential related proteins in LS52 cells was higher than that in LS52 cells. Finally, a cell membrane protein SRSF6 was identified as the target of aptamer LS52, which may serve as a new molecular target for CRC. Therefore, our work provides a systematic novel approach of studying potential biomarkers and a promising tool for cancer diagnosis and therapy.

摘要

结直肠癌(CRC)在全球范围内具有较高的发病率和死亡率。因此,早期诊断和治疗对于提高CRC患者的生存率具有重要意义。在此,我们使用具有相同遗传背景但恶性潜能不同的两种细胞系(LS174T和CL187)作为靶细胞和阴性细胞进行了消减细胞SELEX。两种适配体LS1和LS52对靶细胞LS174T表现出高亲和力,对具有恶性潜能的CRC细胞具有高特异性,并表现出良好的生物学稳定性。此外,我们发现适配体LS1具有促进CRC细胞体外增殖和单克隆形成的作用,并上调了增殖相关蛋白的表达。基于适配体LS52的高亲和力,将LS52用作分子探针可有效区分临床CRC组织与相邻正常组织、CRC转移组织与非转移组织。对磁珠分选细胞中蛋白质表达的分析表明,LS52细胞中恶性潜能相关蛋白的表达高于LS52细胞。最后,鉴定出一种细胞膜蛋白SRSF6为适配体LS52的靶标,其可能作为CRC的新分子靶点。因此,我们的工作提供了一种系统的研究潜在生物标志物的新方法以及一种用于癌症诊断和治疗的有前景的工具。

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