Open AccessQuantitative fluorescent profiling of VEGFR s reveals tumor cell and endothelial cell heterogeneity in breast cancer xenografts
Author(s) -
Imoukhuede Princess. I.,
Popel Aleksander S.
Publication year - 2014
Publication title -
cancer medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.403
H-Index - 53
ISSN - 2045-7634
DOI - 10.1002/cam4.188
Subject(s) - cell , cancer research , endothelial stem cell , biology , receptor , tumor progression , cancer cell , tumor microenvironment , cell growth , cell culture , cancer , microbiology and biotechnology , chemistry , in vitro , tumor cells , biochemistry , genetics
Abstract Plasma membrane‐localized vascular endothelial growth factor receptors ( VEGFR ) play a critical role in transducing VEGF signaling toward pro and antiangiogenic outcomes and quantitative characterization of these receptors is critical toward identifying biomarkers for antiangiogenic therapies, understanding mechanisms of action of antiangiogenic drugs, and advancing predictive computational models. While in vitro analysis of cell surface‐ VEGFR s has been performed, little is known about the levels of cell surface‐ VEGFR on tumor cells. Therefore, we inoculate nude mice with the human triple‐negative breast cancer, MDA ‐ MB ‐231, cell line; isolate human tumor cells and mouse tumor endothelial cells from xenografts; and quantitatively characterize the VEGFR localization on these cells. We observe 15,000 surface‐ VEGFR 1/tumor endothelial cell versus 8200 surface‐ VEGFR 1/tumor endothelial cell at 3 and 6 weeks of tumor growth, respectively; and we quantify 1200–1700 surface‐ VEGFR 2/tumor endothelial cell. The tumor cell levels of VEGFR 1 and VEGFR 2 are relatively constant between 3 and 6 weeks: 2000–2200 surface‐ VEGFR 1/tumor cell and ~1000 surface‐ VEGFR 2/tumor cell. Cell‐by‐cell analysis provides additional insight into tumor heterogeneity by identifying four cellular subpopulations based on size and levels of cell membrane‐localized VEGFR . Furthermore, when these ex vivo data are compared to in vitro data, we observe little to no VEGFR s on MDA ‐ MB ‐231 cells, and the MDA ‐ MB ‐231 VEGFR surface levels are not regulated by a saturating dose of VEGF . Overall, the quantification of these dissimilarities for the first time in tumor provides insight into the balance of modulatory ( VEGFR 1) and proangiogenic ( VEGFR 2) receptors.