The First Affiliated Hospital of Fujian Medical University, the First Clinical Medical College of Fujian Medical University, Fuzhou 350004, PR China.
Analyst. 2018 May 15;143(10):2267-2275. doi: 10.1039/c8an00271a.
Glioma is a cancer derived from transformed glial cells, which are often invasive and display a heterogeneous cell population. Currently, no trustworthy biomarkers for the detection and risk stratification of glioma have been discovered. The objective of the present research was to select DNA aptamers to facilitate early diagnosis and effective therapy of glioma. Using cell-SELEX, three aptamers (WYZ-37, WYZ-41, WYZ-50), which can specifically recognize the molecular differences between target cells T98G and negative cells SVGp12, were identified. The best binding sequences WYZ-41 and WYZ-50 were optimized in length, resulting in aptamer sequences WYZ-41a and WYZ-50a. The Kd values of the aptamers WYZ-41a and WYZ-50a against the target cell line were found to be 1.0 ± 0.2 nM and 2.8 ± 0.6 nM, respectively, which are better than the Kds for full-length aptamers WYZ-41 and WYZ-50. Flow cytometry analysis results show that the aptamers WYZ-41a and WYZ-50a do not influence each other in mutual binding, and that they effectively detect the target even in complex mixtures, such as undiluted fetal bovine serum (FBS) and cerebral spinal fluid (CSF), indicating that aptamers WYZ-41a and WYZ-50a have excellent potential as aptamer pairs to improve the accuracy of glioma diagnosis.
神经胶质瘤是一种源自转化神经胶质细胞的癌症,通常具有侵袭性和异质性细胞群体。目前,尚未发现可靠的生物标志物用于检测和分层胶质瘤的风险。本研究的目的是筛选 DNA 适体,以促进神经胶质瘤的早期诊断和有效治疗。通过细胞 SELEX,鉴定出了三个能够特异性识别靶细胞 T98G 和负细胞 SVGp12 之间分子差异的适体(WYZ-37、WYZ-41、WYZ-50)。对最佳结合序列 WYZ-41 和 WYZ-50 进行了长度优化,得到适体序列 WYZ-41a 和 WYZ-50a。发现适体 WYZ-41a 和 WYZ-50a 对靶细胞系的 Kd 值分别为 1.0 ± 0.2 nM 和 2.8 ± 0.6 nM,优于全长适体 WYZ-41 和 WYZ-50 的 Kd 值。流式细胞术分析结果表明,适体 WYZ-41a 和 WYZ-50a 在相互结合时不会相互影响,并且即使在复杂混合物(如未稀释的胎牛血清(FBS)和脑脊液(CSF))中也能有效地检测到靶标,这表明适体 WYZ-41a 和 WYZ-50a 具有作为适体对的优异潜力,可提高神经胶质瘤诊断的准确性。