Abstract
Anoikis (detachment-induced apoptosis) prevents the survival of cells at inappropriate sites of the body and can therefore act as a barrier to metastasis. In a function-based genome-wide screen, we have previously identified the neurotrophic tyrosine kinase receptor TrkB as a potent suppressor of anoikis. Consistently, activated TrkB oncogenically transforms non-malignant epithelial cells and causes them to invade and produce metastatic tumors in vivo. Overexpression of activated TrkB also results in morphological transformation, resembling epithelial-mesenchymal transition (EMT). E-cadherin, an important EMT regulator, and two E-cadherin repressors, Twist and Snail, are critical for these TrkB functions. As Snail has been shown to induce Zeb1, another E-cadherin repressor, we hypothesized that Zeb1 could be a TrkB target, too. We show here that Zeb1 is required for TrkB-induced EMT in epithelial cells, as RNAi-mediated knockdown of Zeb1 reverted the morphological changes induced by TrkB. Furthermore, Zeb1 is involved in TrkB-induced anoikis resistance, migration and invasion. In vivo, knockdown of Zeb1 strongly reduced TrkB-induced metastasis. Finally, epistasis experiments showed that Zeb1 acts downstream of Twist and Snail. We conclude that Zeb1 is required for several TrkB-induced effects in vitro and in vivo, including metastasis.
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Acknowledgements
We thank M Voetel, S Greven, H Grimminck and all other animal caretakers for their excellent technical help with the in vivo experiments, JY Song and the animal pathology department for help with the pathological analysis and performing IHC stainings, L Brocks and L Oomen for help with the confocal images, F Van Diepen and A Pfauth for fluorescence-activated cell sorting, M Heimerikx, M Nieuwland, J de Ronde, D Sie, A Velds, I de Rink and R Kerkhoven for performing the microarray analysis and ML Yurda for statistical analysis. We thank Dr Nakshatri for providing the Zeb1 plasmid, R Weinberg for the Twist plasmid, A Munoz for the Snail plasmid and K Becker for the Snail antibody. We thank all members of the Peeper laboratory for their valuable input and T Geiger for useful discussions and help with the microarray experiments. This work was supported by grants from MAS and DSP from the Dutch Cancer Society (KWF) and a European Union FP6 grant.
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Smit, M., Peeper, D. Zeb1 is required for TrkB-induced epithelial-mesenchymal transition, anoikis resistance and metastasis. Oncogene 30, 3735–3744 (2011). https://doi.org/10.1038/onc.2011.96
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DOI: https://doi.org/10.1038/onc.2011.96
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