3D hydrogel breast cancer models for studying the effects of hypoxia on epithelial to mesenchymal transition

// Ying Wang 1, 2 , S ameer Mirza 3 , Shaohua Wu 1, 6 , Jiping Zeng 2 , Wen Shi 1, 6 , Hamid Band 3, 4, 5 , Vimla Band 3, 5 and Bin Duan 1, 6, 7, 8 1 Mary & Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, Omaha, NE, USA 2 Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Shandong University, Jinan, China 3 Department of Genetics, Cell Biology and Anatomy University of Nebraska Medical Center, Omaha, NE, USA 4 Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA 5 Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA 6 Division of Cardiology, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, USA 7 Department of Surgery, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA 8 Department of Mechanical and Materials Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA Correspondence to: Bin Duan, email: bin.duan@unmc.edu Vimla Band, email: vband@unmc.edu Keywords: lysyl oxidase; epithelial-mesenchymal transition; hydrogel; hypoxia Received: March 07, 2018     Accepted: July 21, 2018     Published: August 14, 2018 ABSTRACT Solid tumors are 3D assemblies of cancer cells, together with multiple stromal cell types within an extracellular matrix. Yet, the vast majority of cell-based studies to characterize oncogenesis and discovery of new anti-cancer drugs is conducted using conventional 2D monolayer culture systems, where cells are grown on plastic substratum under normoxic environments. In current study, we generated 3D breast cancer cell culture platform consists of photocrosslinkable hydrogels and encapsulated isogenic primary (21PT) and a metastatic (21MT-2) breast cancer cell lines derived from the primary tumor and pleural effusion from the same patient. We demonstrated that hypoxia decreased cellular assembly size and density, and promoted epithelial to mesenchymal transition (EMT) process, without affecting cell viability. Next, we showed hypoxia enhanced breast cancer cell migration, and expression and secretion of lysyl oxidase (LOX), which is copper-dependent amine oxidase and has the primary function to drive the crosslinking of collagen and elastin and is regulated by hypoxia. Furthermore, to recapitulate in vivo situation, we generated breast cancer and lung cells (derived from the same patient) contact model by stacking 3D hydrogel constructs with breast cancer cells onto lung mesenchymal cells (LMC) laden-hydrogel and then showed breast cancer cells migrated towards LMC during hypoxia. Lastly, as a validation of this model for future screen of therapeutic agents, we demonstrated that LOX inhibitor exhibited a significant decrease in breast cancer cell viability, migration, and EMT. Taken together, these results validate the use of hydrogels based models to examine hypoxia related EMT in breast cancer cells.