Characterizing Epithelial-Mesenchymal-Transition (EMT)-Like Behavior in Breast Cancer Cells in a Tissue Engineering Co-Culture Model
Abstract
Tissue engineered 3D cell culture models have shown promise investigating the influence of the tumor microenvironment on cancer cell behavior. Synthetic hydrogels, such as those made from poly(ethylene glycol) (PEG), can be customized with bio-instructive moieties that mimic different parts of the extracellular matrix (ECM) that cells can adhere to through integrins and regions of cadherins that cells can interact with through their own cadherins. The question remains of how the composition of the ECM and neighboring cell interactions impact the transition from an epithelial phenotype to a mesenchymal-like phenotype, termed EMT, which is considered crucial in tumor progression to metastatic cancer. Therefore, in this project, a cell-adhesive peptide that mimics ECM and an cadherin mimetic peptide that mimics mesenchymal N-cadherin were utilized to investigate how cell-ECM interactions through integrin binding and how cell-cell interactions through cadherin binding can direct the emergence of EMT markers in breast cancer. Cytotoxicity studies demonstrated that the peptides allowed high SKBR3 breast cancer cell cytocompatibility but significantly lowered 3T3 fibroblast viability in a 3D environment, indicating that the peptide interactions were changing the behavior of the 3T3’s in a way that did not affect the SKBR3’s. Also, encapsulated as single cells in a gel with a cell-ECM peptide, the 3T3’s began to obtain a spindle-like phenotype, but not the SKBR3’s, showing that the 3T3’s were starting to form focal adhesions and take on a more mesenchymal-like phenotype not taken on by SKBR3’s in the same conditions. When putting the 3T3’s and SKBR3’s in co-culture, the SKBR3’s obtained a spindle-like phenotype when the 3T3’s were in a gel with a cadherin peptide, showing the beginning of an epithelial-mesenchymal transition-like morphology. After running quantitative PCR on said SKBR3’s, it was found that SKBR3’s with 3T3’s in contact with both cell adhesive and cadherin peptide and without either peptides were making significantly more mRNA for Collagen III and vimentin than SKBR3’s in contact with 3T3’s in only a cell-ECM peptide gel, both proteins made as a sign of becoming more mesenchymal-like. Collectively, these results demonstrate that the ability to incorporate mimetic peptides of cell-cell and cell-ECM interaction into PEG hydrogels makes them a powerful platform for studying how the composition of the ECM impacts EMT.
Subject
hydrogeltissue engineering
peptides
biomimetic
cancer
breast cancer
tissue engineering model
co-culture
fibroblasts
EMT
epithelial
mesenchymal
epithelial-mesenchymal transition (EMT)
cell-cell interactions
cell-ECM interactions
tumor microenvironment
tissue model
co-culture model
RGD
HAVDI
PEG
cadherins
integrins
N-cadherin
vimentin
fibronectin
tumor microenvironment
in vitro model
RNA
mRNA
cell adhesion
Citation
Rakoski, Amanda Isabella (2020). Characterizing Epithelial-Mesenchymal-Transition (EMT)-Like Behavior in Breast Cancer Cells in a Tissue Engineering Co-Culture Model. Undergraduate Research Scholars Program. Available electronically from https : / /hdl .handle .net /1969 .1 /196671.
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