Three-dimensional Organotypic Culture Models of Human Hepatocellular Carcinoma.
Three-dimensional Organotypic Culture Models of Human Hepatocellular Carcinoma.
复制标题
人肝细胞癌的三维器官培养模型。
DOI：
10.1038/srep21174
复制
发表时间：
2016-02-16
影响因子：
4.6
通讯作者：
Wang XW
中科院分区：
文献类型：
--
作者：
Takai A;Fako V;Dang H;Forgues M;Yu Z;Budhu A;Wang XW
三维细胞培养方法是可行的体外方法，有助于检查体内存在的癌细胞的生物学特征。在这项研究中，我们证明了在多孔藻酸盐支架中的肝细胞癌（HCC）细胞可以产生类器官样球体，其模拟体内腺上皮的许多特征，例如腺泡形态发生和EpCAM的顶端表达模式，EpCAM是一种在具有干性特征的HCC亚群中高度表达的肝干/祖细胞标志物。我们发现，Wnt/β-连环蛋白信号传导的激活是维持HCC干性的重要途径，是EpCAM+ HCC球状体形成以及腺泡结构维持所必需的。此外，我们证明，与传统的二维（2D）培养相比，培养为球状体的EpCAM+ HCC细胞对TGF/β诱导的上皮-间充质转化更敏感，具有高度的体内致瘤性和转移潜力。此外，EpCAM+球状体中的HCC细胞比2D培养的细胞对化疗剂更具抗性。基于藻酸盐支架的器官型培养系统是一种有前途的、可靠的和简单的系统，其可以被配置成高通量的方式用于鉴定关键信号传导途径和筛选HCC特异性的分子药物靶点。
Three-dimensional cell culture methods are viable in vitro approaches that facilitate the examination of biological features cancer cells present in vivo. In this study, we demonstrate that hepatocellular carcinoma (HCC) cells in porous alginate scaffolds can generate organoid-like spheroids that mimic numerous features of glandular epithelium in vivo, such as acinar morphogenesis and apical expression patterns of EpCAM, a hepatic stem/progenitor cell marker highly expressed in a subset of HCC with stemness features. We show that the activation of Wnt/β-catenin signaling, an essential pathway for maintaining HCC stemness, is required for EpCAM+ HCC spheroid formation as well as the maintenance of the acinous structure. Furthermore, we demonstrate that EpCAM+ HCC cells cultured as spheroids are more sensitive to TGF/β-induced epithelial-mesenchymal transition with highly tumorigenic and metastatic potential in vivo compared to conventional two-dimensional (2D) culture. In addition, HCC cells in EpCAM+ spheroids are more resistant to chemotherapeutic agents than 2D-cultured cells. The alginate scaffold-based organotypic culture system is a promising, reliable, and easy system that can be configured into a high throughput fashion for the identification of critical signaling pathways and screening of molecular drug targets specific for HCC.