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Flow Cytometric FRET Analysis of erbB Receptor Interaction on a Cell‐by‐Cell Basis
Author(s) -
DiermeierDaucher Simone,
Hasmann Max,
Brockhoff Gero
Publication year - 2008
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1430.003
Subject(s) - pertuzumab , trastuzumab , cell growth , microbiology and biotechnology , erbb , chemistry , receptor , signal transduction , cell , skbr3 , monoclonal antibody , cell culture , flow cytometry , intracellular , cancer research , cancer cell , biology , antibody , biochemistry , immunology , cancer , breast cancer , genetics , human breast
Lateral interaction of c‐erbB family receptors resulting in dimer formation is the key event initiating signal transduction. Consequently cross‐activation and intracellular signaling is triggered with immediate impact on cell proliferation, migration, cell survival, and differentiation. In order to elucidate the connection of signal input (receptor activation) and signal output (altered cellular behavior) we dynamically assessed cell proliferation of BT474 and SK‐BR‐3 breast cancer cell lines. We quantitated c‐erbB2 receptor homodimerization upon treatment with the therapeutic monoclonal anti‐c‐erbB2 antibodies trastuzumab (Herceptin®) and pertuzumab by flow cytometric FRET (FCET) measurements on a cell‐by‐cell basis and calculated the extent of antibody‐induced cell cycle exit. The results confirm that trastuzumab does not decrease c‐erbB2 homodimers despite its strong potency to drive c‐erbB2‐overexpressing cells into quiescence. Pertuzumab, however, is able to prevent c‐erbB2 homodimerization and thereby enhance the antiproliferative effect of trastuzumab when administered in combination.