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2D vs 3D – Triple negative breast cancer spheroid formation induces quantitative heterogeneity of VEGF and PDGF receptor profiles and modulates cytosolic phosphorylation
Author(s) -
Oyirifi Ashley,
Joyce Kaitlyn M.,
Helferich William G.,
Imoukhuede Princess I
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.647.30
Subject(s) - platelet derived growth factor receptor , microbiology and biotechnology , phosphorylation , biology , cell , cancer research , receptor , chemistry , growth factor , biochemistry
Micro‐tumor spheres (MTS) have been considered a phenotypic intermediate between 2‐dimensional cell culture and tumor explants from living organisms. However, limited information exists for understanding membrane bound antigens of live cells in MTS. These antigens include several families of transmembrane proteins fundamental to signal transduction, cell communication, and adhesion processes and are thereby identified as key therapeutic targets in malignant cells. This study couples MTS cell culture and a high throughput method for live single cell receptoromic analysis, to compare MTS and monolayer cell surface densities of the PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 tumor angiogenesis regulating receptors on MDB‐MB‐231. Cumulatively, the average density of PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 in MTS is 1.7‐fold higher in tumor only MTS when compared to monolayer cells. When cells are analyzed on a cell‐by‐cell basis, 2‐component Gaussian mixture models predict means of high‐density and low‐density MTS sub‐populations within PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 frequency distributions. The high density to low density means show 3.9‐, 1.6‐, 1.1‐, and 3.2‐fold differences for PDGFRα, PDGFRβ, VEGFR1, and VEGFR2, respectively. Finally, an analysis of MTS intracellular phosphorylation profiles preliminarily reveals attenuated phosphorylation, including reduced activation of the MAPK pathway, in MDA‐MB‐231 MTS simulated by a 20pg/mL VEGFA solution. Together, the current data suggests that MTS may provide a biologically relevant representation of breast cancer cells' VEGFR and PDGFR signaling axes through altering cell surface receptor density and heterogeneity. Our continued work will investigate the effects of varying VEGA concentrations on MTS signal transduction. Support or Funding Information This research was supported by NIEHS grant (T32 ES007326). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .