A microchip filter device incorporating slit arrays and 3-D flow for detection of circulating tumor cells using CAV1-EpCAM conjugated microbeads
A microchip filter device incorporating slit arrays and 3-D flow for detection of circulating tumor cells using CAV1-EpCAM conjugated microbeads
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DOI：
10.1016/j.biomaterials.2014.05.039
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发表时间：
2014-08-01
期刊：
影响因子：
14
通讯作者：
Kim, You-Sun
中科院分区：
文献类型：
--
作者：
Kim, Yeon Jeong;Koo, Gi-Bang;Kim, You-Sun
循环肿瘤细胞（CTC）是罕见的细胞，这些细胞的存在可能表明预后不良和转移的可能性高。尽管CTC具有非常有前途的临床应用，但由于在其分离和鉴定中面临许多技术限制，CTC尚未被彻底研究。目前的CTC检测技术大多采用上皮细胞粘附分子（EpCAM）作为上皮细胞标志物，但越来越多的证据表明，由于上皮细胞向间质细胞转化（EMT），CTC表现出EpCAM的异质性表达。在这项研究中，我们报告了一个微芯片过滤装置，结合狭缝阵列和三维流动，可以分离异质群体的细胞与标记的CTC。为了选择靶点，我们在诱导EMT表型的延长的乳腺球培养条件下培养乳腺癌细胞。在这些条件下，细胞显示出caveolin 1（CAV 1）的上调，但EpCAM表达的下调。所提出的含有CAV 1-EpCAM缀合珠的装置具有几十倍的增加的通量。更重要的是，该平台能够提高转移性乳腺癌患者的捕获率，并获得表达各种EMT标志物的细胞。对这些EMT相关表型的进一步了解将导致检测技术的改进，并可能为开发有效治疗和预防癌症转移的治疗策略提供机会。(C)2014爱思唯尔有限公司版权所有。
Circulating tumor cells (CTCs) are rare cells and the presence of these cells may indicate a poor prognosis and a high potential for metastasis. Despite highly promising clinical applications, CTCs have not been investigated thoroughly, due to many technical limitations faced in their isolation and identification. Current CTC detection techniques mostly take the epithelial marker epithelial cell adhesion molecule (EpCAM), however, accumulating evidence suggests that CTCs show heterogeneous EpCAM expression due to the epithelial-to-mesenchymal transition (EMT). In this study, we report that a microchip filter device incorporating slit arrays and 3-dimensional flow that can separate heterogeneous population of cells with marker for CTCs. To select target we cultured breast cancer cells under prolonged mammosphere culture conditions which induced EMT phenotype. Under these conditions, cells show upregulation of caveolin1 (CAV1) but down-regulation of EpCAM expression. The proposed device which contains CAV1-EpCAM conjugated bead has several tens of times increased throughput. More importantly, this platform enables the enhanced capture yield from metastatic breast cancer patients and obtained cells that expressed various EMT markers. Further understanding of these EMT-related phenotypes will lead to improved detection techniques and may provide an opportunity to develop therapeutic strategies for effective treatment and prevention of cancer metastasis. (C) 2014 Elsevier Ltd. All rights reserved.