Abstract
BackgroundAggressive metastatic breast cancer cells seemingly evade surgical resection and current therapies, leading to colonization in distant organs and tissues and poor patient prognosis. Therefore, high-throughput in vitro tools allowing rapid, accurate, and novel anti-metastatic drug screening are grossly overdue. Conversely, aligned nanofiber constitutes a prominent component of the late-stage breast tumor margin extracellular matrix. This parallel suggests that the use of a synthetic ECM in the form of a nanoscale model could provide a convenient means of testing the migration potentials of cancer cells to achieve a long-term goal of providing clinicians an in vitro platform technology to test the efficacy of novel experimental anti-metastatic compounds.MethodsElectrospinning produces highly aligned, cell-adhesive nanofiber matrices by applying a strong electric field to a polymer-containing solution. The resulting fibrous microstructure and morphology closely resembles in vivo tumor microenvironments suggesting their use in analysis of migratory potentials of metastatic cancer cells. Additionally, a novel interface with a gel-based delivery system creates CXCL12 chemotactic gradients to enhance CXCR4-expressing cell migration.ResultsCellular dispersions of MCF-10A normal mammary epithelial cells or human breast cancer cells (MCF-7 and MDA-MB-231) seeded on randomly-oriented nanofiber exhibited no significant differences in total or net distance traveled as a result of the underlying topography. Cells traveled ~2-5 fold greater distances on aligned fiber. Highly-sensitive MDA-MB-231 cells displayed an 82% increase in net distance traversed in the presence of a CXCL12 gradient. In contrast, MCF-7 cells exhibited only 31% increase and MCF-10A cells showed no statistical difference versus control or vehicle conditions. MCF-10A cells displayed little sensitivity to CXCL12 gradients, while MCF-7 cells displayed early sensitivity when CXCL12 concentrations were higher. MDA-MB-231 cells displayed low relative expression levels of CXCR4, but high sensitivity resulting in 55-fold increase at late time points due to CXCL12 gradient dissipation.ConclusionsThis model could create clinical impact as an in vitro diagnostic tool for rapid assessment of tumor needle biopsies to confirm metastatic tumors, their invasiveness, and allow high-throughput drug screening providing rapid development of personalized therapies.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2407-14-825) contains supplementary material, which is available to authorized users.
Highlights
Aggressive metastatic breast cancer cells seemingly evade surgical resection and current therapies, leading to colonization in distant organs and tissues and poor patient prognosis
QRT-polymerase chain reaction (PCR) for CXCR4 mRNA expression The expression of CXCR4 mRNA was analyzed over a 24 hour period for all three cell lines to determine if exposure to its C-X-C motif chemokine 12 (CXCL12) ligand over time transcriptionally regulates CXCR4 gene expression. 1.5×105 cells were inoculated in 24-well plates in triplicate with or without 100 ng of rhCXCL12 (R&D Systems, Inc; 350-NS-010)
While it is well understood that early detection is the best prevention of metastasis in breast cancer, tumor cells located in the margins following surgical lumpectomy or mastectomy contain populations of dormant cancer stem cells that can result in distant metastases and reduce patient survival [66,67,68,69]
Summary
Aggressive metastatic breast cancer cells seemingly evade surgical resection and current therapies, leading to colonization in distant organs and tissues and poor patient prognosis. Aligned nanofiber constitutes a prominent component of the late-stage breast tumor margin extracellular matrix This parallel suggests that the use of a synthetic ECM in the form of a nanoscale model could provide a convenient means of testing the migration potentials of cancer cells to achieve a long-term goal of providing clinicians an in vitro platform technology to test the efficacy of novel experimental anti-metastatic compounds. Tumor-associated collagen signatures, TACS-3, as described by Conklin et al, is characterized by radially-organized, highly-aligned collagen fibers/bundles located at the tumor-stroma interface potentially providing a topography that enables rapid stromal invasion [25,30] Clinical observation of these aligned collagen bundles oriented perpendicular to the tumor boundary using histological evaluation of patient biopsies correlates to poor prognosis and reduced treatment efficacy [25,31]
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