Purpose Transdifferentiation exists within stromal cells in the tumour microenvironment. Transforming growth factor-β (TGF-β) secreted by tumour-associated fibroblasts (TAFs) affects the differentiation states of epithelial cells, including epithelial-mesenchymal transition (EMT). Evodiamine, a natural drug, can regulate differentiation. However, the specific effects and relative mechanisms of evodiamine remain unknown. Design We used four models to observe the influence of TAF-like CCD-18Co cells on the colon epithelial cell line HCoEpiC: the 3D- and 2D-mono-culture system, Transwell and direct co-culture model. Additionally, we established conditioned medium from CCD-18Co cells. The TGF-β pathway inhibitor LY364947 and evodiamine were added. Morphological changes and classical EMT markers were observed and detected using phase contrast microscopy and immunofluorescence. Cell migration was measured by the wound-healing assay. Western blotting was performed to detect the TGF-β/Smad signalling pathway. Results CCD-18Co cells induced EMT-like changes in the 2D- and 3D-cultured epithelial cell line HCoEpiC, accompanied by high expression of ZEB1 and Snail and the enhancement of migration. Moreover, CCD-18Co-derived conditioned medium caused dysfunction of TGF-β/Smad signalling in EMT. Evodiamine inhibited these EMT-like HCoEpiC and their migration. Additionally, evodiamine down-regulated the expression of ZEB1/Snail and up-regulated the expression of phosphorylated Smad2/3 (pSmad2/3). Evodiamine also increased the ratios of pSmad2/Smad2 and pSmad3/Smad3. Conclusion Based on our observations, evodiamine can reverse the TAF-induced EMT-like phenotype in colon epithelial cells, which may be associated with its mediation of phosphorylated Smad2 and Smad3 expression.
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Bo Liu, PhD, and Cun-jia Zhang, PhD, provided medical writing support.
The work was supported by the Natural Science Foundation of Guangdong Province (2018A030310060) and PhD Initial Foundation of Hospital (20170916).
Conflict of interest
Wan bin Yang declares that he has no conflict of interest. Xiu li Gong declares that she has no conflict of interest. Xiu lian Wang declares that she has no conflict of interest. Chao Huang declares that he has no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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Yang, W., Gong, X., Wang, X. et al. A mediator of phosphorylated Smad2/3, evodiamine, in the reversion of TAF-induced EMT in normal colonic epithelial cells. Invest New Drugs 37, 865–875 (2019). https://doi.org/10.1007/s10637-018-0702-x
- Epithelial-mesenchymal transition
- Tumour-associated fibroblasts
- Tumour microenvironment
- Transforming growth factor-β