HS2ST1‐dependent signaling pathways determine breast cancer cell viability, matrix interactions, and invasive behavior

• Published in: Cancer science Vol. 111; no. 8; pp. 2907 - 2922
• Main Authors: Vijaya Kumar, Archana, Brézillon, Stéphane, Untereiner, Valérie, Sockalingum, Ganesh Dhruvananda, Kumar Katakam, Sampath, Mohamed, Hossam Taha, Kemper, Björn, Greve, Burkhard, Mohr, Benedikt, Ibrahim, Sherif Abdelaziz, Goycoolea, Francisco M., Kiesel, Ludwig, Pavão, Mauro S.G., Motta, Juliana M., Götte, Martin
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01-08-2020; Wiley Open Access; John Wiley and Sons Inc
• Subjects: 2‐O‐sulfotransferase; Antigens, CD - metabolism; Apoptosis; Biochemistry, Molecular Biology; Breast cancer; Breast Neoplasms - pathology; Butadienes - pharmacology; Cadherins - metabolism; Cell adhesion & migration; Cell Movement - drug effects; Cell Survival - drug effects; Cell viability; Cytokines; Cytomegalovirus; Epidermal growth factor; Epidermal growth factor receptors; ErbB Receptors - metabolism; Extracellular matrix; Female; Fibroblast growth factor 2; Fibroblast Growth Factor 2 - metabolism; Fibronectin; Gene Knockdown Techniques; Glycosaminoglycans; Growth factors; Heparan sulfate; Heparan sulfate proteoglycans; Humans; Invasiveness; Laminin; Life Sciences; MAP kinase; MAP Kinase Signaling System - drug effects; MAPK signaling pathway; MCF-7 Cells; Microscopy; Motility; Neoplasm Invasiveness - pathology; Nitriles - pharmacology; Original; Phenotypes; proteoglycan; Proteoglycans; RNA, Small Interfering - metabolism; Signal transduction; siRNA; Sulfates; Sulfotransferase; Sulfotransferases - genetics; Sulfotransferases - metabolism; Tumor cell lines; Wnt protein; Wound healing; 2-O-sulfotransferase; breast cancer; heparan sulfate; MAPK signaling pathway; proteoglycan
• ISSN: 1347-9032; 1349-7006
• DOI: 10.1111/cas.14539

Heparan sulfate proteoglycans (HSPGs) act as signaling co‐receptors by interaction of their sulfated glycosaminoglycan chains with numerous signaling molecules. In breast cancer, the function of heparan sulfate 2‐O‐sulfotransferase (HS2ST1), the enzyme mediating 2‐O‐sulfation of HS, is largely unknown. Hence, a comparative study on the functional consequences of HS2ST1 overexpression and siRNA knockdown was performed in the breast cancer cell lines MCF‐7 and MDA‐MB‐231. HS2ST1 overexpression inhibited Matrigel invasion, while its knockdown reversed the phenotype. Likewise, cell motility and adhesion to fibronectin and laminin were affected by altered HS2ST1 expression. Phosphokinase array screening revealed a general decrease in signaling via multiple pathways. Fluorescent ligand binding studies revealed altered binding of fibroblast growth factor 2 (FGF‐2) to HS2ST1‐expressing cells compared with control cells. HS2ST1‐overexpressing cells showed reduced MAPK signaling responses to FGF‐2, and altered expression of epidermal growth factor receptor (EGFR), E‐cadherin, Wnt‐7a, and Tcf4. The increased viability of HS2ST1‐depleted cells was reduced to control levels by pharmacological MAPK pathway inhibition. Moreover, MAPK inhibitors generated a phenocopy of the HS2ST1‐dependent delay in scratch wound repair. In conclusion, HS2ST1 modulation of breast cancer cell invasiveness is a compound effect of altered E‐cadherin and EGFR expression, leading to altered signaling via MAPK and additional pathways.
Our results suggest that, in breast cancer, high levels of HS2ST1 result in structural changes in heparan sulfate and altered growth factor binding, which leads to attenuated signaling through the MAPK and additional pathways. Reduced signaling and expression of E‐cadherin and epidermal growth factor receptor (EGFR) is associated with reduced viability, adhesion, migration, and invasion of breast cancer cells.