Repression of Wnt-5a impairs DDR1 phosphorylation and modifies adhesion and migration of mammary cells.
The Wnt-5a gene encodes a secreted protein that controls several normal processes during embryogenesis and development of adult tissues by as yet unknown mechanisms. Endogenous expression of Wnt-5a mRNA is known to occur in both mouse and human mammary cell lines. To investigate the biological role of Wnt-5a in the human mammary epithelial cell line HB2, we used an antisense approach to repress endogenous expression of Wnt-5a protein. We also generated a cell population that constitutively overexpresses this protein. We found that overexpression of Wnt-5a protein enhanced cell-to-collagen binding and abolished hepatocyte growth factor-stimulated migration of HB2 transfectants through collagen matrices. Conversely, repression of Wnt-5a protein led to cell scattering, impaired cell-collagen interaction and enhanced cell motility. As we were searching for modified collagen receptors in antisense cells, we discovered that the collagen-binding discoidin domain receptor 1 (DDR1) failed to undergo phosphorylation. In reciprocal experiments, phosphorylation of DDR1 was consistently enabled by expression of Wnt-5a-HA protein in non-Wnt-5a-producing MCF-7 breast cancer cells. Activation of the Wnt/beta-catenin signalling pathway did not influence or mimic the Wnt-5a-mediated effect on DDR1 phosphorylation. These data demonstrate that Wnt-5a protein participates in regulation of adhesion to and migration through collagen and is also a co-factor necessary for collagen-induced activation of DDR1 receptors in mammary epithelial cells.
Jönsson
M
M
Division of Experimental Pathology, Lund University, Malmö University Hospital, SE-205 02 Malmö, Sweden. marzieh.jonsson@pat.mas.lu.se
Andersson
T
T
D001940:Breast / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion; D002455:Cell Division; D002460:Cell Line; D002465:Cell Movement; D048429:Cell Size; D003094:Collagen / Q000378:metabolism; D017077:Culture Media, Conditioned; D003560:Cysts / Q000378:metabolism / Q000473:pathology; D003598:Cytoskeletal Proteins / Q000378:metabolism; D016147:Genes, Tumor Suppressor; D006801:Humans; D039201:Integrin alpha3beta1; D016023:Integrins / Q000378:metabolism; D007700:Kinetics; D009687:Nuclear Proteins; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D016372:RNA, Antisense / Q000235:genetics; D038981:Receptors, Collagen; D015398:Signal Transduction; D015534:Trans-Activators; D014162:Transfection; D014407:Tumor Cells, Cultured; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D051176:beta Catenin
C495270:CTNNB1 protein, human; D017077:Culture Media, Conditioned; D003598:Cytoskeletal Proteins; C403503:FAM107A protein, human; D039201:Integrin alpha3beta1; D016023:Integrins; D009687:Nuclear Proteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D016372:RNA, Antisense; D038981:Receptors, Collagen; D015534:Trans-Activators; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D051176:beta Catenin; D003094:Collagen