title: Acquisition of epithelial-mesenchymal transition and cancer stem-like phenotypes within chitosan-hyaluronan membrane-derived 3D tumor spheroids.
abstract: Cancer drug development has to go through rigorous testing and evaluation processes during pre-clinical in vitro studies. However, the conventional two-dimensional (2D) in vitro culture is often discounted by the insufficiency to present a more typical tumor microenvironment. The multicellular tumor spheroids have been a valuable model to provide more comprehensive assessment of tumor in response to therapeutic strategies. Here, we applied chitosan-hyaluronan (HA) membranes as a platform to promote three-dimensional (3D) tumor spheroid formation. The biological features of tumor spheroids of human non-small cell lung cancer (NSCLC) cells on chitosan-HA membranes were compared to those of 2D cultured cells in vitro. The cells in tumor spheroids cultured on chitosan-HA membranes showed higher levels of stem-like properties and epithelial-mesenchymal transition (EMT) markers, such as NANOG, SOX2, CD44, CD133, N-cadherin, and vimentin, than 2D cultured cells. Moreover, they exhibited enhanced invasive activities and multidrug resistance by the upregulation of MMP2, MMP9, BCRC5, BCL2, MDR1, and ABCG2 as compared with 2D cultured cells. The grafting densities of HA affected the tumor sphere size and mRNA levels of genes on the substrates. These evidences suggest that chitosan-HA membranes may offer a simple and valuable biomaterial platform for rapid generation of tumor spheroids in vitro as well as for further applications in cancer stem cell research and cancer drug screening.
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Yen-Jang
authors.initials: YJ
authors.lastname: Huang
authors.affiliation: Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan. Electronic address: shhsu@mail.nchu.edu.tw.
authors.forename: Shan-Hui
authors.initials: SH
authors.lastname: Hsu
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods; D001672:Biocompatible Materials / Q000138:chemical synthesis*; D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry*; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D008567:Membranes, Artificial; D014411:Neoplastic Stem Cells / Q000473:pathology* / Q000502:physiology*; D010641:Phenotype; D018874:Spheroids, Cellular / Q000473:pathology / Q000502:physiology; D059016:Tumor Microenvironment / Q000502:physiology
chemical_list: D001672:Biocompatible Materials; D008567:Membranes, Artificial; D006820:Hyaluronic Acid; D048271:Chitosan
keywords: Cancer stem cell; Epithelial–mesenchymal transition; Hyaluronan; Multidrug resistance; Tumor spheroids