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Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab

Abstract

Three-dimensional (3D) cell culture techniques are frequently used to model alterations in tissue architecture critically important for tumor development. Here, we report on a detailed comparison of a spheroid model of human epidermal growth factor receptor (HER2) overexpressing cancer cells with the traditional monolayer culture. In 2D culture, HER2 and HER3 form heterodimers, whereas in multicellular spheroids HER2 homodimers are formed. These homodimers localize in membrane rafts, resulting in enhanced inhibition of the proliferation of cancer cells with trastuzumab (Herceptin), a monoclonal antibody specifically targeting HER2. Within the tumor spheroids, HER2 homodimerization leads to enhanced activation of HER2 and results in a switch in signaling pathways from phosphoinositide 3-kinase (PI3K) to mitogen-activated protein kinase (MAPK). Diminished PI3K signaling is accompanied by the activation of the integrin β4/Rac1/PAK 2 signaling cascade. We propose that the described 3D culture system may better reflect some in vivo aspects of HER signaling and can be used to further improve the understanding of the molecular mechanisms of trastuzumab action. Furthermore, the described human multicellular tumor spheroids may allow identification of new targets for the treatment of HER2-positive breast cancer patients who currently benefit suboptimally from trastuzumab treatment.

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Acknowledgements

We thank Leopoldo Luistro for providing the SKOV-3 cell line. KPL-4 cells were kindly provided by Professor J Kurebayashi, Kawasaki Medical School, Kurashiki, Okayama, Japan. We are grateful to Olaf Mundigl and Heike Seul for their assistance with confocal microscopy, and Ute Baer for technical assistance with gene expression profiling. We gratefully acknowledge Manfred Kubbies for fruitful discussions and expert advice, Max Hasmann and Mark Sliwkowski for helpful discussions, Stefan Ries for critical review of the paper and Lynda Chang for medical writing support.

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Correspondence to C H Ries.

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Pickl, M., Ries, C. Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab. Oncogene 28, 461–468 (2009). https://doi.org/10.1038/onc.2008.394

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Keywords

  • HER2 signaling
  • membrane raft
  • trastuzumab
  • 3D cell culture

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