Hwang Ji Yeon, Kim Sang Tae, Han Ho-Seong, Kim Kyunggon, Han Jin Soo
Preclinical Research Center, Biomedical Research Institute, Seoul National University Bundang Hospital, 82, Gumi-ro 173beon-gil, Bundang-gu, Seongnam-si, Gyeonggi-do 13620, Korea.
The Institute for the 3Rs, College of Veterinary Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Korea.
Sensors (Basel). 2016 Nov 23;16(11):1909. doi: 10.3390/s16111909.
Fluorescence detecting of exogenous EpCAM (epithelial cell adhesion molecule) or muc1 (mucin1) expression correlated to cancer metastasis using nanoparticles provides pivotal information on CTC (circulating tumor cell) occurrence in a noninvasive tool. In this study, we study a new skill to detect extracellular EpCAM/muc1 using quantum dot-based aptamer beacon (QD-EpCAM/muc1 ALB (aptamer linker beacon). The QD-EpCAM/muc1 ALB was designed using QDs (quantum dots) and probe. The EpCAM/muc1-targeting aptamer contains a Ep-CAM/muc1 binding sequence and BHQ1 (black hole quencher 1) or BHQ2 (black hole quencher2). In the absence of target EpCAM/muc1, the QD-EpCAM/muc1 ALB forms a partial duplex loop-like aptamer beacon and remained in quenched state because the BHQ1/2 quenches the fluorescence signal-on of the QD-EpCAM/muc1 ALB. The binding of EpCAM/muc1 of CTC to the EpCAM/muc1 binding aptamer sequence of the EpCAM/muc1-targeting oligonucleotide triggered the dissociation of the BHQ1/2 quencher and subsequent signal-on of a green/red fluorescence signal. Furthermore, acute inflammation was stimulated by trigger such as caerulein in vivo, which resulted in increased fluorescent signal of the cy5.5-EpCAM/muc1 ALB during cancer metastasis due to exogenous expression of EpCAM/muc1 in Panc02-implanted mouse model.
使用纳米颗粒对与癌症转移相关的外源性上皮细胞黏附分子(EpCAM)或黏蛋白1(MUC1)表达进行荧光检测,可为非侵入性工具中循环肿瘤细胞(CTC)的出现提供关键信息。在本研究中,我们研究了一种使用基于量子点的适体信标(QD-EpCAM/MUC1 ALB,适体连接体信标)检测细胞外EpCAM/MUC1的新技术。QD-EpCAM/MUC1 ALB是使用量子点(QDs)和探针设计的。靶向EpCAM/MUC1的适体包含一个Ep-CAM/MUC1结合序列和黑洞猝灭剂1(BHQ1)或黑洞猝灭剂2(BHQ2)。在没有目标EpCAM/MUC1的情况下,QD-EpCAM/MUC1 ALB形成部分双链环状适体信标,并保持淬灭状态,因为BHQ1/2淬灭了QD-EpCAM/MUC1 ALB的荧光信号开启。CTC的EpCAM/MUC1与靶向EpCAM/MUC1的寡核苷酸的EpCAM/MUC1结合适体序列结合,触发了BHQ1/2猝灭剂的解离以及随后绿色/红色荧光信号的开启。此外,体内通过如蛙皮素等触发因素刺激急性炎症,这导致在Panc02植入小鼠模型中癌症转移期间,由于EpCAM/MUC1的外源性表达,cy5.5-EpCAM/MUC1 ALB的荧光信号增加。