Lipid tethering of breast tumor cells enables real-time imaging of free-floating cell dynamics and drug response.
Lipid tethering of breast tumor cells enables real-time imaging of free-floating cell dynamics and drug response.
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DOI：
10.18632/oncotarget.7251
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发表时间：
2016-03-01
期刊：
影响因子：
--
通讯作者：
Jewell CM
中科院分区：
文献类型：
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作者：
Chakrabarti KR;Andorko JI;Whipple RA;Zhang P;Sooklal EL;Martin SS;Jewell CM
位于癌症患者血液中的自由漂浮肿瘤细胞，称为循环肿瘤细胞（CTC），已成为研究转移的关键目标。然而，体外研究肿瘤细胞自由漂浮行为的有效策略一直是限制对 CTC 功能特性理解的主要障碍。细胞外基质（ECM）脱离后，乳腺肿瘤细胞形成基于微管蛋白的突起，称为微触手（McTN），在肿瘤细胞的聚集和重新附着中发挥作用，以提高其转移效率。在这项研究中，我们设计了一种策略来空间固定 ECM 分离的肿瘤细胞，同时保持其自由漂浮的特性。我们使用沉积在微流体基底上的聚电解质多层来防止肿瘤细胞粘附，并添加脂质部分通过与细胞膜的相互作用将肿瘤细胞束缚在这些表面上。该涂层保持光学透明，可以捕获活的自由漂浮细胞上的 McTN 的高分辨率图像和视频。此外，我们还表明，束缚可以实时分析单个肿瘤细胞的 McTN 动态以及对微管蛋白靶向药物的反应。对分离的肿瘤细胞进行成像的能力可以极大地增强我们对 CTC 的理解，从而更好地重现它们在转移过程中遇到的微环境。
Free-floating tumor cells located in the blood of cancer patients, known as circulating tumor cells (CTCs), have become key targets for studying metastasis. However, effective strategies to study the free-floating behavior of tumor cells in vitro have been a major barrier limiting the understanding of the functional properties of CTCs. Upon extracellular-matrix (ECM) detachment, breast tumor cells form tubulin-based protrusions known as microtentacles (McTNs) that play a role in the aggregation and re-attachment of tumor cells to increase their metastatic efficiency. In this study, we have designed a strategy to spatially immobilize ECM-detached tumor cells while maintaining their free-floating character. We use polyelectrolyte multilayers deposited on microfluidic substrates to prevent tumor cell adhesion and the addition of lipid moieties to tether tumor cells to these surfaces through interactions with the cell membranes. This coating remains optically clear, allowing capture of high-resolution images and videos of McTNs on viable free-floating cells. In addition, we show that tethering allows for the real-time analysis of McTN dynamics on individual tumor cells and in response to tubulin-targeting drugs. The ability to image detached tumor cells can vastly enhance our understanding of CTCs under conditions that better recapitulate the microenvironments they encounter during metastasis.