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Mesothelial cells interact with tumor cells for the formation of ovarian cancer multicellular spheroids in peritoneal effusions

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Abstract

Epithelial ovarian cancer (EOC) dissemination is primarily mediated by the shedding of tumor cells from the primary site into ascites where they form multicellular spheroids that rapidly lead to peritoneal carcinomatosis. While the clinical importance and fundamental role of multicellular spheroids in EOC is increasingly appreciated, the mechanisms that regulate their formation and dictate their cellular composition remain poorly characterized. To investigate these important questions, we characterized spheroids isolated from ascites of women with EOC. We found that in these spheroids, a core of mesothelial cells was encased in a shell of tumor cells. Analysis further revealed that EOC spheroids are dynamic structures of proliferating, non-proliferating and hypoxic regions. To recapitulate these in vivo findings, we developed a three-dimensional co-culture model of primary EOC and mesothelial cells. Our analysis indicated that, compared to the OVCAR3 cell line, primary EOC cells isolated from ascites as well as mesothelial cells formed compact spheroids. Analysis of heterotypic spheroid microarchitecture revealed a structure that grossly resembles the structure of spheroids isolated from ascites. Cells that formed compact spheroids had elevated expression of β1 integrin and low expression of E-cadherin. Addition of β1 integrin blocking antibody or siRNA-mediated downregulation of β1 integrin resulted in reduced tightness of the spheroids. Interestingly, the loss of MUC16 and E-cadherin expression resulted in the formation of more compact spheroids. Therefore, our findings support the heterotypic nature of spheroids from malignant EOC ascites. In addition, our data describe an unusual link between E-cadherin expression and less compact spheroids. Our data also emphasize the role of MUC16 and β1 integrin in EOC spheroid formation.

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Acknowledgments

This work was supported by an internal Grant from the Université de Sherbrooke, by the Centre d’excellence en Inflammation-Cancer de l’Université de Sherbrooke and by the “Programme d’aide de financement interne” of the Centre de Recherche du Centre Hospitalier Universitaire de Sherbrooke. We wish to thank the Banque de tissus et de données du Réseau de Recherche en Cancer du Fond de Recherche du Québec en Santé (FRQS), affiliated to the Canadian Tumor Repository Network (CTRNet) for providing the ascites samples and primary cells.

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Authors and Affiliations

  1. Département de Microbiologie et Infectiologie, Université de Sherbrooke, 3001, 12ième Avenue Nord, Sherbrooke, QC, J1H 5N4, Canada

    Isabelle Matte, Clara Major Legault, Claudine Rancourt & Alain Piché

  2. Département de Pathologie, Université de Sherbrooke, 3001, 12ième Avenue Nord, Sherbrooke, QC, J1H 5N4, Canada

    Perrine Garde-Granger & Claude Laplante

  3. Département de Chirurgie, Service d’Obstétrique-Gynécologie, Faculté de Médecine, Université de Sherbrooke, 3001, 12ième Avenue Nord, Sherbrooke, QC, J1H 5N4, Canada

    Paul Bessette

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  1. Isabelle Matte
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Correspondence to Alain Piché.

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Matte, I., Legault, C.M., Garde-Granger, P. et al. Mesothelial cells interact with tumor cells for the formation of ovarian cancer multicellular spheroids in peritoneal effusions. Clin Exp Metastasis 33, 839–852 (2016). https://doi.org/10.1007/s10585-016-9821-y

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