title: Cell surface heparan sulfate proteoglycans control adhesion and invasion of breast carcinoma cells.
abstract: BACKGROUND
  Cell surface proteoglycans interact with numerous regulators of cell behavior through their glycosaminoglycan chains. The syndecan family of transmembrane proteoglycans are virtually ubiquitous cell surface receptors that are implicated in the progression of some tumors, including breast carcinoma. This may derive from their regulation of cell adhesion, but roles for specific syndecans are unresolved.
  
  
  METHODS
  The MDA-MB231 human breast carcinoma cell line was exposed to exogenous glycosaminoglycans and changes in cell behavior monitored by western blotting, immunocytochemistry, invasion and collagen degradation assays. Selected receptors including PAR-1 and syndecans were depleted by siRNA treatments to assess cell morphology and behavior. Immunohistochemistry for syndecan-2 and its interacting partner, caveolin-2 was performed on human breast tumor tissue arrays. Two-tailed paired t-test and one-way ANOVA with Tukey's post-hoc test were used in the analysis of data.
  
  
  RESULTS
  MDA-MB231 cells were shown to be highly sensitive to exogenous heparan sulfate or heparin, promoting increased spreading, focal adhesion and adherens junction formation with concomitantly reduced invasion and matrix degradation. The molecular basis for this effect was revealed to have two components. First, thrombin inhibition contributed to enhanced cell adhesion and reduced invasion. Second, a specific loss of cell surface syndecan-2 was noted. The ensuing junction formation was dependent on syndecan-4, whose role in promoting actin cytoskeletal organization is known. Syndecan-2 interacts with, and may regulate, caveolin-2. Depletion of either molecule had the same adhesion-promoting influence, along with reduced invasion, confirming a role for this complex in maintaining the invasive phenotype of mammary carcinoma cells. Finally, both syndecan-2 and caveolin-2 were upregulated in tissue arrays from breast cancer patients compared to normal mammary tissue. Moreover their expression levels were correlated in triple negative breast cancers.
  
  
  CONCLUSIONS
  Cell surface proteoglycans, notably syndecan-2, may be important regulators of breast carcinoma progression through regulation of cytoskeleton, cell adhesion and invasion.
authors.affiliation: Department of Biomedical Sciences and Biotech Research & Innovation Center, University of Copenhagen, Biocenter, Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark. hooi_ching.lim@med.lu.se.|Current address: Stem Cell Center, Lund University, Lund, Sweden. hooi_ching.lim@med.lu.se.
authors.forename: Hooi Ching
authors.initials: HC
authors.lastname: Lim
authors.affiliation: Department of Biomedical Sciences and Biotech Research & Innovation Center, University of Copenhagen, Biocenter, Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark. hinke.multhaupt@bric.ku.dk.
authors.forename: Hinke A B
authors.initials: HA
authors.lastname: Multhaupt
authors.affiliation: Department of Biomedical Sciences and Biotech Research & Innovation Center, University of Copenhagen, Biocenter, Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark. john.couchman@bric.ku.dk.
authors.forename: John R
authors.initials: JR
authors.lastname: Couchman
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D051243:Caveolin 2 / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010641:Phenotype; D044463:Receptor, PAR-1 / Q000378:metabolism; D015398:Signal Transduction; D053669:Syndecan-2 / Q000235:genetics / Q000378:metabolism; D053671:Syndecan-4 / Q000235:genetics / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism
chemical_list: D051243:Caveolin 2; D019812:Heparan Sulfate Proteoglycans; D044463:Receptor, PAR-1; D053671:Syndecan-4; D053669:Syndecan-2; D006493:Heparin; D013917:Thrombin; D053511:Matrix Metalloproteinase 14