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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McCluggage WG. Malignant biphasic uterine tumours: carcinosarcomas or metaplastic carcinomas? J Clin Pathol. 2002;55:321–5.
McCluggage WG. Uterine carcinosarcomas (malignant mixed mullerian tumors) are metaplastic carcinomas. Int J Gynecol Cancer. 2002;12:687–90.
D’Angelo E, Prat J. Uterine sarcomas: a review. Gynecol Oncol. 2010;23:694–702.
Thompson L, Chang B, Barsky SH. Monoclonal origins of malignant mixed tumors (carcinosarcoma). Evidence for a divergent histogenesis. Am J Surg Pathol. 1996;20:277–85.
Gupta GP, Massague J. Cancer metastasis: building a framework. Cell. 2006;127:679–95.
Casas E, Kim J, Bendesky A, Ohno-Machado L, Wolfe CJ, Yang J. Snail2 is an essential mediator of twist-induced epithelial mesenchymal transition and metastasis. Cancer Res. 2011;71:245–54.
Savagner P, Yamada KM, Thiery JP. The zinc-finger protein slug causes desmosome dissociation, an initial and necessary step for growth factor-induced epithelial-mesenchymal transition. J Cell Biol. 1997;137:1403–19.
Batlle E, Sancho E, Franci C, Dominguez D, Monfar M, Baulida J. et al. The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol. 2000;2:84–9.
Bolos V, Peinado H, Perez-Moreno MA, Fraga MF, Esteller M, Cano A. The transcription factor Slug represses E-cadherin expression and induces epithelial to mesenchymal transitions: a comparison with Snail and E47 repressors. J Cell Sci. 2003;116:499–511.
Li J, Zhou BP. Activation of β-catenin and Akt pathways by Twist are critical for the maintenance of EMT associated cancer stem cell like characters. BMC Cancer. 2011;11:49.
Thiery JP. Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002;2:442–54.
Sarrio D, Rodriguez-Pinilla SM, Hardisson D, Cano A, Moerno-Bueno G, Palacios J. Epithelial-mesenchymal transition in breast cancer relates to the basal-like phenotype. Cancer Res. 2006;68:989–97.
Castilla MA, Moreno-Bueno G, Romero-Perez L, Van De Vijver K, Biscuola M, Lopez-Garcia MA, et al. Micro-RNA signature of the epithelial-mesenchymal transition in endometrial carcinosarcoma. J Pathol. 2011;223:72–80.
Reya T, Morrison SJ, Clarke MF, Weissman IL. Stem cells, cancer, and cancer stem cells. Nature. 2001;414:105–11.
Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133:704–15.
Griorian M, Andresen S, Tulchinsky E, Kriajevska M, Carlberg C, Kruse C, et al. Tumor suppressor p53 protein is a new target for the metastasis-associated Mts1/S100A4 protein: functional consequences of their interaction. J Biol Chem. 2001;276:22699–708.
Li Z-H, Bresnick AR. The S100A4 metastasis factor regulates cellular motility via a direct interaction with myosin-IIA. Cancer Res. 2006;66:5173–80.
Semov A, Moreno MJ, Onichtchenko A, Abulrob A, Ball M, Ekiel I, et al. Metastasis-associated protein S100A4 induces angiogenesis through interaction with Annexin II and accelerated plasmin formation. J Biol Chem. 2005;280:20833–41.
Chow K-H, Park HJ, George J, Yamamoto K, Gallup AD, Graber JH, et al. S100A4 is a biomarker and regulator of glioma stem cells that is critical for mesenchymal transition in glioblastoma. Cancer Res. 2017;77:5360–73.
Akiya M, Yamazaki M, Matsumoto T, Kawashima Y, Oguri Y, Kajita S, et al. Identification of LEFTY as a molecular marker for ovarian clear cell carcinoma. Oncotarget. 2017;8:63646–64.
Saegusa M, Hashimura M, Kuwata T, Hamano M, Okayasu I. Crosstalk between NF-κB/p65 and β-catenin/TCF4/p300 signalling pathways through alterations in GSK-3β expression during trans-differentiation of endometrial carcinoma cells. J Pathol. 2007;213:35–45.
Inoue H, Takahashi H, Hashimura M, Eshima K, Akiya M, Matsumoto T et al. Cooperation of Sox4 with β-catenin/p300 complex in transcriptional regulation of the Slug gene during divergent sarcomatous differentiation in uterine carcinosarcoma. BMC Cancer. 2016;16:53.
Saegusa M, Hashimura M, Kuwata T. Sox4 functions as a positive regulator of β-catenin signaling through upregulation of TCF4 during morular differentiation of endometrial carcinomas. Lab Invest. 2012;92:511–21.
Inoue H, Hashimura M, Akiya M, Chiba R, Saegusa M. Functional role of ALK-related signal cascades on modulation of epithelial-mesenchymal transition and apoptosis in uterine carcinosarcoma. Mol Cancer. 2017;16:37.
Saegusa M, Hashimura M, Kuwata T, Hamano M, Okayasu I. β-Catenin simultaneously induces activation of the p53-p21WAF1 pathway and overexpression of cyclin D1 during squamous differentiation of endometrial carcinoma cells. Am J Pathol. 2004;164:1739–49.
Kawashima Y, Takahashi N, Satoh M, Saito T, Kado S, Nomura F et al. Enhanced recovery of lyophilized peptides in shotgun proteomics by using an LC-ESI-MC compatible surfactant. Proteomics. 2013;13:751–5.
Kawashima Y, Kodera Y, Singh A, Matsumoto M, Matsumoto H. Efficient extraction of proteins from formalin-fixed paraffin-embedded tissues requires higher concentration of tris(hydroxymethyl)aminomethane. Clin Proteomics. 2014;11:4.
Zaino R, Carinelli SG, Ellenson LH, Eng C, Katabuchi H, Konishi I, et al. Tumours of the uterine corpus. In: Kurman RJ, Carcangiu ML, Herrington CS, Young RH, editors. WHO classification of tumours of female reproductive organs. Lyon: IARC; 2014. p. 121–54.
Fei F, Qu J, Li C, Wang X, Li Y, Zhang S. Role of metastasis-induced protein S100A4 in human non-tumor pathophysiologies. Cell Biosci. 2017;7:64.
Mazzucchelli L. Protein S100A4: too long overlooked by pathologists? Am J Pathol. 2002;160:7–13.
Helfman DM, Kim EJ, Lukanidin E, Grigorian M. The metastasis associated protein A100A4: role of tumour progression and metastasis. Br J Cancer. 2005;92:1955–8.
Boye K, Malandsmo GM. S100A4 and metastasis: a small actor playing many roles. Am J Pathol. 2010;176:528–35.
Straight AF, Cheung A, Limouze J, Chen I, Westwood NJ, Sellers JR, et al. Dissecting temporal and spatial control of cytokinesis with a myosin II inhibitor. Science. 2003;299:1743–7.
Kas SM, de Ruiter JR, Schipper K, Annunziato S, Schut E, Klarenbeek S, et al. Insertional mutagenesis identifies drivers of a novel oncogenic pathway in invasive lobular breast carcinoma. Nat Genet. 2017;49:1219–30.
Cohn MA, Hjeimso I, Wu L-C, Guldberg P, Lukanidin EM, Tulchinsky EM. Characterization of Sp1, AP-1, CBF and KRC binding sites and minisatellite DNA as functional elements of the metastasis-associated mts1/S100A4 gene intronic enhancer. Nucleic Acids Res. 2001;29:3335–46.
Freudlsperger C, Bian Y, Wise SC, Burnett J, Coupar J, Yang X et al. Van Waes C. TGF-β and NF-κB signal pathway cross-talk is mediated through TAK1 and SMAD7 in a subset of head and neck cancers. Oncogene. 2013;32:1549–59.
Egeland EV, Boye K, Pettersen SJ, Haugen MH, Oyjord T, Malerod L et al. Enrichment of nuclear S100A4 during G2/M in colorectal cancer cells: possible association with cyclin B1 and centrosomes. Clin Exp Metastasis. 2015;32:755–67.
Giese A, Loo MA, Tran N, Haskett D, Coons SW, Berens ME. Dichotomy of astrocytoma migration and proliferation. Int J Cancer. 1996;67:275–82.
Giese A, Bjerkvig R, Berens ME, Westphal M. Cost of migration: invasion of malignant glioma and implications for treatment. J Clin Oncol. 2003;21:1624–36.
Merzak A, McCrea S, Koocheckpour S, Pilkington GJ. Control of human glioma cell growth, migration and invasion in vitro by transforming growth factor beta 1. Br J Cancer. 1994;70:199–203.
Kriajevska M, Tarabykina S, Bronstein I, Maitland N, Lomonosov M, Hansen K et al. Metastasis-associated Mts1 (S100A$) protein modulates protein kinase C phosphorylation of the heavy chain of nonmuscle myosin. J Biol Chem. 1998;273:9852–6.
Li Z-H, Spektor A, Varlamova O, Bresnick AR. Mts1 regulates the assemble of nonmuscle myosin-IIA. Biochemistry. 2003;42:14258–66.
Pecci A, Ma X, Savoia A, Adelstein RS. MYH9: structure, functions and role of non-muscle myosin IIA in human disease. Gene. 2018;664:152–67.
Lo J-F, Yu C-C, Chiou S-H, Huang C-Y, Jan C-I, Lin S-C et al. The epithelial-mesenchymal transition mediator S100A4 maintains cancer-initiating cells in head and neck cancers. Cancer Res. 2011;71:1912–23.
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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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