EGF-induced redistribution of erbB2 on breast tumor cells: Flow and image cytometric energy transfer measurements

Péter Nagy, László Bene, Margit Balázs, William C. Hyun, Steven J. Lockett, Nancy Y. Chiang, Frederick Waldman, Burt G. Feuerstein, Sándor Damjanovich, János Szöllosi

Research output: Contribution to journalArticle

45 Scopus citations

Abstract

erbB2, a member of the epidermal growth factor (EGF) receptor-type tyrosine kinase receptor family, is overexpressed in breast carcinomas with poor prognosis. We examined the cell surface association of this receptor with itself and with other cell surface proteins by the Forster-type fluorescence resonance energy transfer using whole antibodies and Fab fragments. We found that erbB2 molecules homoassociate in unstimulated SK- BR-3, BT474 and BT474-M (a metastatic version of the parent BT474 line) breast tumor cells, and that the interaction was enhanced by EGF treatment in suspensions of SK-BR-3 and BT474-M cells. BT474 cells (with low EGF receptor expression) and attached SK-BR-3 cells do not respond to EGF. Image microscopic energy transfer measurements found considerable pixel-by-pixel heterogeneity in the homoassociation state of erbB2. In accordance with the EGF-induced redistribution of erbB2, EGF receptor was found to be in close proximity to erbB2 in FRET measurements. B labeling different epitopes on erbB2 and the lipid bilayer, we were able to prepare an epitope map of erbB2 molecule. Our data suggest the existence of dynamic cell surface patterns of erbB2 and point to functions fulfilled by these molecular complexes.

Original languageEnglish (US)
Pages (from-to)120-131
Number of pages12
JournalCytometry
Volume32
Issue number2
DOIs
StatePublished - Jun 1 1998
Externally publishedYes

Keywords

  • Breast cancer
  • EGF receptor
  • Energy transfer
  • Homoassociation
  • erbB2

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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    Nagy, P., Bene, L., Balázs, M., Hyun, W. C., Lockett, S. J., Chiang, N. Y., Waldman, F., Feuerstein, B. G., Damjanovich, S., & Szöllosi, J. (1998). EGF-induced redistribution of erbB2 on breast tumor cells: Flow and image cytometric energy transfer measurements. Cytometry, 32(2), 120-131. https://doi.org/10.1002/(SICI)1097-0320(19980601)32:2<120::AID-CYTO7>3.0.CO;2-P