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A mediator of phosphorylated Smad2/3, evodiamine, in the reversion of TAF-induced EMT in normal colonic epithelial cells

  • Wanbin Yang
  • Xiuli Gong
  • Xiulian Wang
  • Chao Huang
PRECLINICAL STUDIES
  • 19 Downloads

Summary

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.

Keywords

Epithelial-mesenchymal transition Tumour-associated fibroblasts Tumour microenvironment Transdifferentiation Transforming growth factor-β Evodiamine 

Notes

Acknowledgements

Bo Liu, PhD, and Cun-jia Zhang, PhD, provided medical writing support.

Funding

The work was supported by the Natural Science Foundation of Guangdong Province (2018A030310060) and PhD Initial Foundation of Hospital (20170916).

Compliance with ethical standards

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.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

For this type of study, formal consent is not required.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Tropical DiseasesGuangzhou University of Chinese MedicineGuangzhouPeople’s Republic of China
  2. 2.Department of Spleen and Stomach DiseasesSecond Hospital of Traditional Chinese Medicine of GuangdongGuangzhouPeople’s Republic of China
  3. 3.Department of Traditional Chinese Medicine, Affiliated Bao’an Hospital of Traditional Chinese Medicine of ShenzhenGuangzhou University of Chinese MedicineShenzhenPeople’s Republic of China
  4. 4.Central Laboratory, Affiliated Bao’an Hospital of ShenzhenSouthern Medical UniversityShenzhenPeople’s Republic of China

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