Flow cytometric FRET analysis of erbB receptor interaction on a cell-by-cell basis
Flow cytometric FRET analysis of erbB receptor interaction on a cell-by-cell basis
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DOI：
10.1196/annals.1430.003
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发表时间：
2008-01-01
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影响因子：
--
通讯作者：
Brockhoff, Gero
中科院分区：
文献类型：
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作者：
Diermeier-Daucher, Simone;Hasmann, Max;Brockhoff, Gero
c-erbB家族受体的横向相互作用导致二聚体的形成是启动信号转导的关键事件。因此，交叉激活和细胞内信号被触发，对细胞增殖、迁移、细胞存活和分化产生直接影响。为了阐明信号输入（受体激活）和信号输出（细胞行为改变）之间的联系，我们动态评估了BT474和SK-BR-3乳腺癌细胞系的细胞增殖。我们在使用治疗性单克隆抗c-erbB2抗体曲妥珠单抗（赫赛汀(R)）和帕妥珠单抗治疗后，通过流式细胞术FRET （FCET）测量逐细胞定量c-erbB2受体同二聚化，并计算抗体诱导的细胞周期退出的程度。结果证实，曲妥珠单抗不降低c-erbB2同型二聚体，尽管其强大的效力驱动c-erbB2过表达的细胞进入静止状态。然而，帕妥珠单抗能够阻止c-erbB2二聚体化，从而增强曲妥珠单抗联合使用时的抗增殖作用。
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(R)) 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.