a Under normal conditions the detachment of anchorage-dependant cells from their extracellular matrix typically induces programmed cell death which is mediated through a pathway referred to as anoikis.However, a resistance to anoikis in cancer enables the migration of cells from the primary tumour and the establishment of aggressive metastatic disease. Although cancer cell aggregation is known to be an important mechanism within anoikis resistance, research into the underlying mechanisms that govern this process remain problematic as commercially available tissue culture material can only sustain 2D monolayer or 3D aggregate/spheroid cultures. This necessitates the development of a system that can accommodate for cancer cell aggregation-disaggregation as a single dynamic process, without the disruption of passaging cells between alternate substrates. This study describes a procedure for modifying tissue culture polystyrene (TCP) to produce a fluoro-silica (FS) surface which preferentially promotes the deposition of a distinct profile of proteins/factors from serum which mediate the transient aggregation of human breast cancer cell lines. This modified surface therefore provides an experimental platform for better understanding cancer cell aggregation-disaggregation events in vitro, and their influence on the establishment of metastatic disease in patients with cancer.
OPCT-1
is a heterogeneous prostate cancer cell line derived from
primary (rather than metastatic) disease which contains epithelial,
mesenchymal, and CD44high/CD24low cancer stem
cell (CSC) subpopulations and from which we have previously generated
and characterized stable mesenchymal (P4B6B) and epithelial (P5B3)
cell subpopulations. In this contribution, we explore the effect of
tissue culture surface chemistry (standard tissue culture plastic
(TCP) and a fluoroalkylsilica (FS) culture surface with inherently
low surface energy) on the phenotype and adherent capacity of mesenchymal
and epithelial cell populations. We demonstrate that OPCT-1 cells
adherent to FS surfaces comprise both epithelial- and mesenchymal-like
populations; a mesenchymal subpopulation derived from OPCT1 (P4B6B)
poorly adheres to FS and formed spheroids, whereas an epithelial subpopulation
derived from OPCT1 (P5B3) forms an adherent monolayer. In contrast,
P4B6B cells do adhere to FS when cocultured with P5B3 cells. Taken
together, these findings demonstrate that EMT/cell differentiation
status dictates cell adhesive capacity and provide a novel insight
into the relationship between epithelial and mesenchymal cell populations
in metastasis. Importantly, the differences in adherence capacity
between P4B6B and P5B3 are not apparent using standard TCP-based culture,
thereby highlighting the value of using alternative culture surfaces
for studying cell surface interaction/adhesion phenomena and interrogating
mechanisms involved in adhesion and detachment of metastatic tumor
cells.
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