Abstract
Progression of colorectal cancer (CRC) involves spatial and temporal occurrences of epithelial–mesenchymal transition (EMT), whereby tumour cells acquire a more invasive and metastatic phenotype. Subsequently, the disseminated mesenchymal tumour cells must undergo a reverse transition (mesenchymal–epithelial transition, MET) at the site of metastases, as most metastases recapitulate the pathology of their corresponding primary tumours. Importantly, initiation of tumour growth at the secondary site is the rate-limiting step in metastasis. However, investigation of this dynamic reversible EMT and MET that underpins CRC morphogenesis has been hindered by a lack of suitable in vitro models. To this end, we have established a unique in vitro model of CRC morphogenesis, which we term LIM1863-Mph (morphogenetic). LIM1863-Mph cells spontaneously undergo cyclic transitions between two-dimensional monolayer (migratory, mesenchymal) and three-dimensional sphere (carcinoid, epithelial) states. Using RNAi, we demonstrate that FZD7 is necessary for MET of the monolayer cells as loss of FZD7 results in the persistence of a mesenchymal state (increased SNAI2/decreased E-cadherin). Moreover, FZD7 is also required for migration of the LIM1863-Mph monolayer cells. During development, FZD7 orchestrates either migratory or epithelialization events depending on the context. Our findings strongly implicate similar functional diversity for FZD7 during CRC morphogenesis.
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Acknowledgements
We thank BL Eckhardt and RL Anderson for advice with RNAi; NJ Waterhouse and S Asquith for assistance with live cell imaging; M Trivett for assistance with FZD7 immunohistochemistry; EW Thompson, M Bills, RH Whitehead, SA Currie for helpful discussion. This work was supported, in part, by funding from the Cancer Council of Victoria (EV). RGR is supported by an NHMRC fellowship.
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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Vincan, E., Darcy, P., Farrelly, C. et al. Frizzled-7 dictates three-dimensional organization of colorectal cancer cell carcinoids. Oncogene 26, 2340–2352 (2007). https://doi.org/10.1038/sj.onc.1210026
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DOI: https://doi.org/10.1038/sj.onc.1210026
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