Detection of Molecular Biomarkers involved in the Induction of EMT, migration and invasion, and drug resistance in breast cancer cells in 3D confinement
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Date
2018-09-17Author
Hill, Tamara Nicole
0000-0001-8129-3172
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Standard-of-care regimes for cancer treatment have become more effective and sophisticated over the last
decade due to research developments which facilitate specific targeting of treatments to certain cancers,
and the ability to provide specialized care to cancer patients. However, cancer treatments overall still fall
short of preventing spread of disease from the primary tumor to secondary sites where they settle and form
new more aggressive tumors. It is this stage of disease progression which results in the majority of cancer
deaths. The long-term goal of this research is to develop a system for the effective high throughput screening
of chemotherapeutic drugs (and possibly in combination with radiation therapy) that target migrating cancer
cells, for metastatic cancer treatment. In Aim 1 we developed a polydimethylsiloxane (PDMS) microfluidic
device equipped with microchannels for the study of proteome expression changes that are likely induced
through Epithelial-to-mesenchymal (EMT) transition, a result of migration through confined threedimensional
(3D) space. Quantification of protein expression differences between cells that migrated, and
cells that grew in a classical two-dimensional (2D) environment revealed a pattern of molecular changes
associated with EMT which highly invasive MDA-MB-231 breast cancer cells likely underwent while
migrating through the microchannels. While examining these differences between cells in the two different
conditions, we were inspired to develop an Engineered Basal Lamina (EBL) for 2D culture of breast and
lung cancer cells. Coating of cell culture well plates with the EBL revealed a significant difference in both
protein expression and chemotherapeutic response from cells grown on the classic polystyrene coating. We
further sought to examine in Aim 3, the induction of dedifferentiation of breast cancer cells upon migration
through confined space, and the molecular properties associated with this process that render these cells
completely unresponsive to Doxorubicin (DOX), a potent breast cancer drug whose mode of action is DNA
intercalation and inhibition of macromolecular biosynthesis. We sought to uncover the genetic and
epigenetic activities of the cancer cells upon collection from the microchannels and re-culture in a 2D environment, as well as changes associated with multiple and extended migration events. This would likely uncover possible therapeutic targets for cells which have migrated to secondary sites.