Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation.

Zeinab Dehghani-Ghobadi,Ehsan Arefian,Ghamartaj Hossein,Shahrzad Sheikh Hasani

doi:10.3390/cells11020237

Zeinab Dehghani-Ghobadi, Ehsan Arefian + Show 2 more

Open Access

https://doi.org/10.3390/cells11020237

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Journal: Cells	Publication Date: Jan 11, 2022
Citations: 10	License type: CC BY 4.0

Affiliation: University of Tehran, Imam Khomeini Hospital

Abstract

In this paper, we investigate whether Wnt5A is associated with the TGF-β1/Smad2/3 and Hippo-YAP1/TAZ-TEAD pathways, implicated in epithelial to mesenchymal transition (EMT) in epithelial ovarian cancer. We used 3D and 2D cultures of human epithelial ovarian cancer cell lines SKOV-3, OVCAR-3, CAOV-4, and different subtypes of human serous ovarian cancer compared to normal ovary specimens. Wnt5A showed a positive correlation with TAZ and TGFβ1 in high- and low-grade serous ovarian cancer specimens compared to borderline serous and normal ovaries. Silencing Wnt5A by siRNAs significantly decreased Smad2/3 activation and YAP1 expression and nuclear shuttling in ovarian cancer (OvCa) cells. Furthermore, Wnt5A was required for TGFβ1-induced cell migration and invasion. In addition, inhibition of YAP1 transcriptional activity by Verteporfin (VP) altered OvCa cell migration and invasion through decreased Wnt5A expression and inhibition of Smad2/3 activation, which was reverted in the presence of exogenous Wnt5A. We found that the activation of TGFβ1 and YAP1 nuclear shuttling was promoted by Wnt5A-induced integrin alpha v. Lastly, Wnt5A was implicated in activating human primary omental mesothelial cells and subsequent invasion of ovarian cancer cells. Together, we propose that Wnt5A could be a critical mediator of EMT-associated pathways.

Highlights

Epithelial ovarian cancer (EOC) is the most common type of ovarian cancer, in which widespread peritoneal dissemination is the main route of metastasis and ascites
We found that Wnt5A is required for TGFβ1 activation and Smad2/3 activation through enhanced Yes-associated protein-1 (YAP1) nuclear shuttling and subsequent up-regulation of integrin αv
We found that the expression levels of WNT5A, ROR1, ROR2, TGFB1, TGFB2, TGFBR1, TGFBR2, YAP1, transcriptional co-activator with PDZ-binding motif (TAZ), CCN1, and CCN2 were significantly different between normal and cancerous groups (p < 2−16)

Summary

Introduction

Epithelial ovarian cancer (EOC) is the most common type of ovarian cancer, in which widespread peritoneal dissemination is the main route of metastasis and ascites. EOC development and metastasis are associated with fibrosis [2], one of the driving forces behind epithelial-to-mesenchymal transition (EMT). Our published studies and other reports showed that Wnt5A exhibits a tumor-promoting effect and could be associated with EMT in EOC progression [6,7,8]. Ovarian cancer (OvCa) metastasis requires mesothelial to mesenchymal transition (MMT) of the peritoneum. These “activated mesothelial cells” promote metastasis by supporting tumor cell adhesion, invasion, and proliferation [9]. A xenograft mouse model showed that host-derived Wnt5A promoted ovarian tumor cell adhesion to peritoneal mesothelial cells as well as migration and invasion, leading to colonization of peritoneal explants [10]. How Wnt5A promotes EMT in OvCa at the molecular level remains obscure

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