微流控多层纸芯片组织阵列用于乳腺癌上皮-间质转化行为研究

The epithelial to mesenchymal transition (EMT), which endows cancer cells with migratory and invasive properties, plays crucial roles in tumor metastasis. A cell culture platform that allows continuous monitoring the dynamic EMT process is of grest interest, as it promises probing the interactions between cancer cells and microenvironmental factors. For this reason, the current study proposes a microfluidic chip platform with embedded multilayer-paper supported tissue array. Different types of constituent elements of the breast cancer tissue, including vascular endothelial cell, fibroblast and breast cancer cell, are seeded on wax-printed papers, respectively, and the breast cancer tissue array is to be constructed by stacking these paper layer by layer. By mimicking tissue structure and cell-microenvironment communication, such a microfluidic chip is able to set up a microenvironment closely to that of a breast cancer in vivo. In this work, breast cancer cell lines with different metastasis potential are to be used to construct the artificial breast cancer tissue. We will assess biofidelity of these chip-based tumor tissues and then stimulate these tissues with different types of EMT related microenvironmental factors. After cell cultures, the multilayer-paper supported tissue will be destacked, thereby allowing detection of cells on different layers. The viability, distribution, morphology, phenotype, genetic structure and gene expression of cells on each layer are to be detected at successive time-points. Through this study, we expect to understand the cell-microenvironment interaction during EMT from a viewpoint of both temporal and spatial.

上皮-间质转化（EMT）赋予癌细胞迁移和侵袭的能力，因而是造成肿瘤转移的重要因素。目前EMT研究面临的一个技术难题是缺少监测肿瘤内部癌细胞阶段性演变的手段，因而无法得知癌细胞与微环境相互作用的动态过程。本研究拟发展一种基于微流控原理的多层纸芯片细胞培养平台。将构成乳腺癌组织的多种细胞成份分别种植于多张图形化纸芯片上，通过叠加构建包含细胞-细胞及细胞-间质相互作用的多层纸芯片乳腺癌组织阵列。通过仿真组织结构及细胞-微环境作用形式，在芯片上细致模拟肿瘤微环境。实验使用不同转移性能的乳腺癌细胞株构建组织阵列，系统评价芯片乳腺癌组织的仿真效果继而模拟体内微环境因素对其施加不同类型刺激。培养不同时间后，拆解多层纸芯片检测乳腺癌组织内各层面细胞生存、分布、形态、表型、基因结构和表达变化，以获取癌细胞EMT演变的动态信息。我们期望藉此研究从时间和空间角度深入了解EMT中乳腺癌细胞与微环境相互作用过程。

上皮-间质转化（EMT）赋予癌细胞迁移和侵袭的能力，因而是造成肿瘤转移的重要因素。目前EMT 研究面临的一个技术难题是缺少监测肿瘤内部癌细胞阶段性演变的手段，因而无法得知癌细胞与微环境相互作用的动态过程。本研究发展了一种基于微流控原理的多层纸芯片细胞培养平台。将构成乳腺癌组织的多种细胞成份分别种植于多张图形化纸芯片上，通过叠加构建包含细胞-细胞及细胞-间质相互作用的多层纸芯片乳腺癌组织阵列。通过仿真组织结构及细胞-微环境作用形式，在芯片上细致模拟肿瘤微环境。实验使用不同转移性能的乳腺癌细胞株构建组织阵列，系统评价芯片乳腺癌组织的仿真效果继而模拟体内微环境因素对其施加不同类型刺激。培养不同时间后，拆解多层纸芯片检测乳腺癌组织内各层面细胞生存、分布、形态、表型、基因结构和表达变化，以获取癌细胞EMT 演变的动态信息。我们的研究结果显示，多层纸芯片乳腺癌组织微阵列构建简便、仿真度高，是肿瘤微环境研究和抗肿瘤药物反应测试的有利工具。