A homogeneous, Anti-dsDNA antibody-based assay for multicolor detection of cancer stem cell transcription factors
A homogeneous, Anti-dsDNA antibody-based assay for multicolor detection of cancer stem cell transcription factors
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基于抗 dsDNA 抗体的均质检测，用于癌症干细胞转录因子的多色检测
DOI：
10.1016/j.aca.2018.04.060
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发表时间：
2018
影响因子：
6.2
通讯作者：
Liu Ping
中科院分区：
文献类型：
--
作者：
Ma Jiehua;Shi Hai;Zhang Meiling;Li Chao;Xiang Yang;Liu Ping
肿瘤干细胞（cancer stem cells，CSC）是维持肿瘤生长、转移和复发的重要细胞。肿瘤干细胞转录因子（Oct 4、Sox 2和Nanog）的高表达是一个有价值的预后因子，提示肿瘤复发和转移的风险较高。因此，开发一种简便、经济的方法来多重检测这些转录因子（TF）是非常必要的。在这项工作中，我们提出了一个通用的均相检测这些TF的抗dsDNA抗体修饰的Fe 3 O 4磁铁矿纳米粒子（aadMNP）的基础上。在分析物的存在下，染料标记的dsDNA被特异性TF结合，这将抑制dsDNA和aadMNP之间的相互作用，产生更高的荧光，可以为免疫传感过程提供信号读出。方法的线性范围为3 ~ 1200 ng/mL，检出限为0.035 ng/mL。此外，我们已经提出了用于灵敏、选择性和快速检测细胞提取物中Oct 4、Sox 2和Nanog的测定，以及突变结合序列的结合亲和力的分析。本研究为临床肿瘤干细胞的检测提供了潜在的应用价值，也为TF结合位点的核苷酸多态性研究提供了新的契机。
Cancer stem cells (CSCs) are responsible for maintaining tumor growth, metastasis and recurrence. The high expression of cancer stem cell transcription factors (Oct4, Sox2 and Nanog) is a valuable prognostic factor, suggesting a higher risk of tumor recurrence and metastasis. So, the development of a convenient and cost-effective method for multiplex assay of these transcription factors (TFs) is highly required. In this work, we have proposed a universal homogeneous assay for multicolor detection of these TFs based on anti-dsDNA antibody-decorated Fe3O4magnetite nanoparticles (aadMNPs). In the presence of analytes, the dye-labeled dsDNAs are bound by specific TFs, which will inhibit the interactions between the dsDNAs and aadMNPs, generating higher fluorescence that may provide signal readout for the immunosensing process. By using the proposed method, Oct4 can be determined in a linear range from 3 to 1200 ng/mL with a detection limit of 0.035 ng/mL. Furthermore, we have presented assays for the sensitive, selective and rapid detection of Oct4, Sox2 and Nanog in cell extract, as well as the analysis of binding affinity of the mutated binding sequences. This work may provide potential applications in clinical CSCs detections, and may open new opportunity for the study of nucleotide polymorphisms in TF binding sites.