Maimaitiyiming Yasen, Yang Chang, Wang Yun, Hussain Liaqat, Naranmandura Hua
Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, People's Republic of China.
Department of Toxicology, School of Medicine and Public Health, Zhejiang University, Hangzhou, People's Republic of China.
Biotechnol Appl Biochem. 2019 May;66(3):412-418. doi: 10.1002/bab.1737. Epub 2019 Feb 21.
Aptamers are short, single-stranded nucleic acid (DNA or RNA) oligonucleotides that can be obtained by a technique called systematic evolution of ligands by exponential enrichment (SELEX) in vitro. Due to superior properties such as small size, high binding affinity, and stability, they are considered to be feasible tools for diagnosis and treatment of disease. In the current study, we attempted to screen a high-affinity DNA aptamer to selectively target the colorectal carcinoma Caco-2 cells by using cell-based SELEX approach. After 14 consecutive rounds of selection, aptamer ApC1 was identified. Confocal microscopy results revealed that ApC1 could rapidly internalize into Caco-2 cells but not HEK 293 cells. Moreover, it showed high specificity to Caco-2 cells rather than other cell lines such as 293T, HeLa, MCF-7, HL-60, and NB4. Collectively, our results demonstrated that aptamer ApC1 has high specificity to colorectal carcinoma Caco-2 cells, which could be further applied for targeted therapy of colorectal cancer in future studies.
适体是短的单链核酸(DNA或RNA)寡核苷酸,可通过一种称为指数富集配体系统进化(SELEX)的体外技术获得。由于其具有诸如尺寸小、结合亲和力高和稳定性好等优异特性,它们被认为是疾病诊断和治疗的可行工具。在本研究中,我们尝试通过基于细胞的SELEX方法筛选一种高亲和力DNA适体,以选择性靶向结肠直肠癌Caco-2细胞。经过连续14轮筛选,鉴定出适体ApC1。共聚焦显微镜结果显示,ApC1可快速内化进入Caco-2细胞,但不能进入HEK 293细胞。此外,它对Caco-2细胞表现出高特异性,而对其他细胞系如293T、HeLa、MCF-7、HL-60和NB4则不然。总体而言,我们的结果表明适体ApC1对结肠直肠癌Caco-2细胞具有高特异性,在未来研究中可进一步应用于结直肠癌的靶向治疗。