S100A4/non-muscle myosin II signaling regulates epithelial-mesenchymal transition and stemness in uterine carcinosarcoma


Uterine carcinosarcoma (UCS) represents a true example of cancer associated with epithelial-mesenchymal transition (EMT), which exhibits cancer stem cell (CSC)-like traits. Although S100A4 is an inducer of EMT, little is known about its involvement in UCS tumorigenesis. Herein, we focused on the functional role of S100A4 during development of UCS. Expression of S100A4 and molecules associated with its function were also examined in 35 UCS cases. In endometrial carcinoma cell lines, S100A4 promoter activity and mRNA levels were significantly increased by the transfection of NF-κB/p65, independent of a putative κB-binding site in the promoter. Cells stably overexpressing S100A4 showed enhancement of CSC properties, along with decreased cell proliferation and acceleration of cell migration. These phenotypes were abrogated in S100A4-knockdown cells. A combination of S100A4 antibody-mediated co-immunoprecipitation and shotgun proteomics analysis revealed that S100A4 strongly interacted with non-muscle myosin II (NMII) heavy chains, including myosin 9 and myosin 14. Specific inhibition of NMII by blebbistatin phenocopied S100A4 overexpression and induced a fibroblast-like morphology. In clinical samples, S100A4 score was significantly higher in sarcomatous as compared with carcinomatous components of UCS, and was positively correlated with ALDH1, Slug, and vimentin scores, and inversely with Ki-67 labeling indices. These findings suggest that an S100A4/NMII-related signaling cascade may contribute to the establishment and maintenance of EMT/CSC properties, along with changes in cell proliferation and migration capability. These events may be initiated in carcinomatous components in UCS and lead to divergent sarcomatous differentiation.

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Fig. 1: Regulation of S100A4 expression in Em Ca cells.
Fig. 2: Relationship of S100A4 expression with cell proliferation and migration in Em Ca cells.
Fig. 3: Relationship between S100A4 expression and CSC properties in Em Ca cells.
Fig. 4: S100A4 frequently interacts with NMII in Em Ca cells.
Fig. 5: Inhibition of NMII by blebbistatin alters cell morphology, proliferation, and migration in Em Ca cells.
Fig. 6: Inhibition of NMII by blebbistatin enhances CSC properties in Em Ca cells.
Fig. 7: IHC findings in serial sections of UCS tissues.
Fig. 8: Schematic representation of the role of an S100A4/NMII-driven signaling cascade as progenitor for divergent sarcomatous differentiation in UCS.


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We thank Dr Robert Adelstein for the generous gift of the CMV-GFP-NMHC II-A plasmid used in this study. This study was supported by a grant from JSPS KAKENHI Grant Number 17K08703.

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MT and MS carried out the majority of the experiments, analyzed the data, and wrote the manuscript. They were helped by MH, YO, and AY. RK, MT, and YK were involved in the study design and data collection. All authors reviewed and approved the final manuscript.

Correspondence to Makoto Saegusa.

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Tochimoto, M., Oguri, Y., Hashimura, M. et al. S100A4/non-muscle myosin II signaling regulates epithelial-mesenchymal transition and stemness in uterine carcinosarcoma. Lab Invest (2019). https://doi.org/10.1038/s41374-019-0359-x

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