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MDA-MB-231 breast cancer cell viability, motility and matrix adhesion are regulated by a complex interplay of heparan sulfate, chondroitin−/dermatan sulfate and hyaluronan biosynthesis

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Abstract

Proteoglycans and glycosaminoglycans modulate numerous cellular processes relevant to tumour progression, including cell proliferation, cell-matrix interactions, cell motility and invasive growth. Among the glycosaminoglycans with a well-documented role in tumour progression are heparan sulphate, chondroitin/dermatan sulphate and hyaluronic acid/hyaluronan. While the mode of biosynthesis differs for sulphated glycosaminoglycans, which are synthesised in the ER and Golgi compartments, and hyaluronan, which is synthesized at the plasma membrane, these polysaccharides partially compete for common substrates. In this study, we employed a siRNA knockdown approach for heparan sulphate (EXT1) and heparan/chondroitin/dermatan sulphate-biosynthetic enzymes (β4GalT7) in the aggressive human breast cancer cell line MDA-MB-231 to study the impact on cell behaviour and hyaluronan biosynthesis. Knockdown of β4GalT7 expression resulted in a decrease in cell viability, motility and adhesion to fibronectin, while these parameters were unchanged in EXT1-silenced cells. Importantly, these changes were associated with a decreased expression of syndecan-1, decreased signalling response to HGF and an increase in the synthesis of hyaluronan, due to an upregulation of the hyaluronan synthases HAS2 and HAS3. Interestingly, EXT1-depleted cells showed a downregulation of the UDP-sugar transporter SLC35D1, whereas SLC35D2 was downregulated in β4GalT7-depleted cells, indicating an intricate regulatory network that connects all glycosaminoglycans synthesis. The results of our in vitro study suggest that a modulation of breast cancer cell behaviour via interference with heparan sulphate biosynthesis may result in a compensatory upregulation of hyaluronan biosynthesis. These findings have important implications for the development of glycosaminoglycan-targeted therapeutic approaches for malignant diseases.

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Acknowledgments

We would like to thank Birgit Pers, Paola Moretto and Sara Deleonibus for expert technical assistance. Funding was provided by German Academic Exchange Service DAAD Grants A/08/15601 (to MV), and EU H2020 RISE-MSCA Project grant number 645756 (GLYCANC) (to MG and AP). The authors acknowledge the Ph.D. School in Biological and Medical Sciences for Ilaria Caon and Elena Caravà fellowships.

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Authors and Affiliations

  1. Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy

    Manuela Viola, Evgenia Karousou, Ilaria Caon, Elena Caravà, Davide Vigetti, Giancarlo De Luca & Alberto Passi

  2. Department of Gynaecology and Obstetrics, Muenster University Hospital, Muenster, Germany

    Kathrin Brüggemann & Martin Götte

  3. Department of Radiotherapy-Radiooncology, University Hospital Muenster, Muenster, Germany

    Burkhard Greve

  4. Institute of Physiology II, University of Muenster, Muenster, Germany

    Christian Stock

  5. Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany

    Christian Stock

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  1. Manuela Viola
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  2. Kathrin Brüggemann
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  9. Giancarlo De Luca
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Correspondence to Manuela Viola or Alberto Passi.

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Viola, M., Brüggemann, K., Karousou, E. et al. MDA-MB-231 breast cancer cell viability, motility and matrix adhesion are regulated by a complex interplay of heparan sulfate, chondroitin−/dermatan sulfate and hyaluronan biosynthesis. Glycoconj J 34, 411–420 (2017). https://doi.org/10.1007/s10719-016-9735-6

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  • DOI: https://doi.org/10.1007/s10719-016-9735-6

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