title: 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.
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.
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy. manuela.viola@uninsubria.it.
authors.forename: Manuela
authors.initials: M
authors.lastname: Viola
authors.affiliation: Department of Gynaecology and Obstetrics, Muenster University Hospital, Muenster, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Brüggemann
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Karousou
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Caon
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Caravà
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Vigetti
authors.affiliation: Department of Radiotherapy-Radiooncology, University Hospital Muenster, Muenster, Germany.
authors.forename: Burkhard
authors.initials: B
authors.lastname: Greve
authors.affiliation: Institute of Physiology II, University of Muenster, Muenster, Germany.|Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Stock
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: De Luca
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, Varese, Italy. alberto.passi@uninsubria.it.
authors.forename: Alberto
authors.identifier: 0000-0003-3328-3089
authors.initials: A
authors.lastname: Passi
authors.affiliation: Department of Gynaecology and Obstetrics, Muenster University Hospital, Muenster, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Götte
mesh_terms: D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D002809:Chondroitin Sulfates / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D003871:Dermatan Sulfate / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006497:Heparan Sulfate / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D000076002:Hyaluronan Synthases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006820:Hyaluronic Acid / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D042361:Mammary Glands, Human / Q000378:metabolism / Q000473:pathology; D009004:Monosaccharide Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D009237:N-Acetyllactosamine Synthase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D033704:Nucleotide Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D000099058:Exostosin 1; D000099237:UDP-Galactose Translocators
chemical_list: D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D006497:Heparan Sulfate; D000076002:Hyaluronan Synthases; D006820:Hyaluronic Acid; D007527:Isoenzymes; D009004:Monosaccharide Transport Proteins; D017351:N-Acetylglucosaminyltransferases; D009237:N-Acetyllactosamine Synthase; D033704:Nucleotide Transport Proteins; D034741:RNA, Small Interfering; D000099058:Exostosin 1; D000099237:UDP-Galactose Translocators; C425807:SLC35D1 protein, human; C487823:SLC35D2 protein, human; C069524:dermatan sulfate chondroitin sulfate; C523794:HAS1 protein, human
keywords: EXT1; Galactosyltransferase I; Glycosaminoglycans; Solute carriers; UDP-sugars; β4GalT7