Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function
Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function
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DOI：
10.1073/pnas.1210182109
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发表时间：
2012-08-21
影响因子：
11.1
通讯作者：
Kamm, Roger D.
中科院分区：
文献类型：
--
作者：
Zervantonakis, Ioannis K.;Hughes-Alford, Shannon K.;Kamm, Roger D.
肿瘤细胞进入血流是癌症转移的关键步骤。尽管在体内观察肿瘤细胞的运动已经取得了重大进展，但肿瘤细胞血管内渗的潜在机制仍然很大程度上是未知的。我们开发了一种基于微流体的分析方法，可以在三维中重建肿瘤-血管界面，从而实现高分辨率、实时成像和精确量化内皮屏障功能。研究的目的是验证这一假设，即癌细胞的血管内皮细胞受到来自相互作用的细胞和细胞与巨噬细胞相互作用的生化因素的调节。我们开发了一种方法来测量空间分辨的内皮通透性，并表明通过分泌肿瘤坏死因子α与巨噬细胞进行信号传递会导致内皮屏障受损。在这些条件下，活体成像证实了血管内插入率的增加。为了进一步研究肿瘤-内皮(TC-EC)信号，我们使用了高侵袭性的纤维肉瘤细胞，并量化了在控制和干扰(与肿瘤坏死因子α)屏障条件下的肿瘤细胞迁移动力学和TC-EC相互作用。我们发现内皮屏障受损与更多的TC-EC相互作用和更快的动力学有关，这与我们的肿瘤血管内皮细胞转移结果一致。综上所述，我们的结果提供了证据，证明内皮细胞对肿瘤细胞的血管内流构成了屏障，这种屏障可以受到肿瘤微环境中存在的因素的调节。
Entry of tumor cells into the blood stream is a critical step in cancer metastasis. Although significant progress has been made in visualizing tumor cell motility in vivo, the underlying mechanism of cancer cell intravasation remains largely unknown. We developed a microfluidic-based assay to recreate the tumor-vascular interface in three-dimensions, allowing for high resolution, real-time imaging, and precise quantification of endothelial barrier function. Studies are aimed at testing the hypothesis that carcinoma cell intravasation is regulated by biochemical factors from the interacting cells and cellular interactions with macrophages. We developed a method to measure spatially resolved endothelial permeability and show that signaling with macrophages via secretion of tumor necrosis factor alpha results in endothelial barrier impairment. Under these conditions intravasation rates were increased as validated with live imaging. To further investigate tumor-endothelial (TC-EC) signaling, we used highly invasive fibrosarcoma cells and quantified tumor cell migration dynamics and TC-EC interactions under control and perturbed (with tumor necrosis factor alpha) barrier conditions. We found that endothelial barrier impairment was associated with a higher number and faster dynamics of TC-EC interactions, in agreement with our carcinoma intravasation results. Taken together our results provide evidence that the endothelium poses a barrier to tumor cell intravasation that can be regulated by factors present in the tumor microenvironment.