Bioengineered three-dimensional co-culture of cancer cells and endothelial cells: A model system for dual analysis of tumor growth and angiogenesis
Bioengineered three-dimensional co-culture of cancer cells and endothelial cells: A model system for dual analysis of tumor growth and angiogenesis
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DOI：
10.1002/bit.26297
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发表时间：
2017-08-01
影响因子：
3.8
通讯作者：
Luo, Kathy Qian
中科院分区：
文献类型：
--
作者：
Chiew, Geraldine Giap Ying;Wei, Na;Luo, Kathy Qian
血管生成标志着良性局部肿瘤转变为危及生命的疾病。许多体外实验都是在二维（2D）平台上进行的，然而，研究肿瘤和内皮细胞（ECs）在三维（3D）平台上生长的行为的研究有限。本研究提供肿瘤细胞与内皮细胞的三维共培养球体，以研究内皮细胞与肿瘤细胞的相互作用。在与HepG2肝细胞共培养（HCC）细胞的三维共培养中，ECs分化形成小管网络。在模型系统中加入血管生成因子或血管生成抑制剂可以增强或抑制3D模型中的内皮分化，从而可以研究HCC发展中利用的细胞信号通路。3D模型显示出与肝癌异种移植相似的蛋白表达水平，并且在3D模型中表现出Akt/mTor等必需信号蛋白的上调，这在2D模型中没有反映出来。在三维共培养模型中，利用荧光蛋白和内皮细胞中基于荧光共振能量转移（FRET）的caspase-3传感器实时检测细胞凋亡，分析索拉非尼、舒尼替尼和阿西替尼等几种抗血管生成药物的作用。通过FRET传感器可以很容易地区分药物在3D模型中抑制血管生成的凋亡能力，并且通过3D共培养模型可以一步实现抗血管生成和抗肿瘤药物的双重筛选。综上所述，我们构建了一个三维共培养模型，在其中实现了一个具有缺氧核心和药物真正梯度渗透的HCC肿瘤微环境，用于药物筛选和利用小型HCC肿瘤的体外研究。Biotechnol。Bioeng。2017;114: 1865 - 1877。(c) 2017 Wiley期刊公司
Angiogenesis marks the transformation of a benign local tumor into a life-threatening disease. Many in vitro assays are available on two-dimensional (2D) platforms, however, limited research has been conducted to investigate the behavior of tumors and endothelial cells (ECs) grown on three-dimensional (3D) platforms. This study provides a 3D co-culture spheroid of tumor cells with ECs to study the interplay between ECs and tumor cells. In a 3D co-culture with HepG2 hepatocellular carcinoma (HCC) cells, ECs differentiate to form tubule networks when in co-culture. Addition of angiogenic factors or angiogenesis inhibitors to the model system enhanced or inhibited endothelial differentiation in the 3D model, enabling investigations of the cellular signaling pathways utilized in HCC development. The 3D model demonstrated similar protein expression levels as a HCC xenograft, as well as exhibited upregulation of essential signaling proteins such as Akt/mTor in the 3D model, which is not reflected in the 2D model. The effects of several anti-angiogenic agents, such as sorafenib, sunitinib, and axitinib were analyzed in the 3D co-culture model by utilizing fluorescent proteins and a fluorescence resonance energy transfer (FRET)-based caspase-3 sensor in the ECs, which can detect apoptosis in real time. The apoptotic capability of a drug to inhibit angiogenesis in the 3D model can be easily distinguished via the FRET sensor, and dual screening of anti-angiogenesis and anti-tumor drugs can be achieved in a single step via the 3D co-culture model. In summary, a 3D co-culture model is constructed, where a HCC tumor microenvironment with a hypoxic core and true gradient penetration of drugs is achieved for drug screening purposes and in vitro studies utilizing a small HCC tumor. Biotechnol. Bioeng. 2017;114: 1865-1877. (c) 2017 Wiley Periodicals, Inc.