title: Tenascin C induces epithelial-mesenchymal transition-like change accompanied by SRC activation and focal adhesion kinase phosphorylation in human breast cancer cells.
abstract: Tenascin C (TNC) is an extracellular matrix glycoprotein up-regulated in solid tumors. Higher TNC expression is shown in invading fronts of breast cancer, which correlates with poorer patient outcome. We examined whether TNC induces epithelial-mesenchymal transition (EMT) in breast cancer. Immunohistochemical analysis of invasive ductal carcinomas showed that TNC deposition was frequent in stroma with scattered cancer cells in peripheral margins of tumors. The addition of TNC to the medium of the MCF-7 breast cancer cells caused EMT-like change and delocalization of E-cadherin and β-catenin from cell-cell contact. Although amounts of E-cadherin and β-catenin were not changed after EMT in total lysates, they were increased in the Triton X-100-soluble fractions, indicating movement from the membrane into the cytosol. In wound healing assay, cells were scattered from wound edges and showed faster migration after TNC treatment. The EMT phenotype was correlated with SRC activation through phosphorylation at Y418 and phosphorylation of focal adhesion kinase (FAK) at Y861 and Y925 of SRC substrate sites. These phosphorylated proteins colocalized with αv integrin-positive adhesion plaques. A neutralizing antibody against αv or a SRC kinase inhibitor blocked EMT. TNC could induce EMT-like change showing loss of intercellular adhesion and enhanced migration in breast cancer cells, associated with FAK phosphorylation by SRC; this may be responsible for the observed promotion of TNC in breast cancer invasion.
authors.affiliation: Department of Pathology and Matrix Biology, Graduate School of Medicine, Mie University, Mie, Japan.
authors.forename: Keiki
authors.initials: K
authors.lastname: Nagaharu
authors.forename: Xinhui
authors.initials: X
authors.lastname: Zhang
authors.forename: Toshimichi
authors.initials: T
authors.lastname: Yoshida
authors.forename: Daisuke
authors.initials: D
authors.lastname: Katoh
authors.forename: Noriko
authors.initials: N
authors.lastname: Hanamura
authors.forename: Yuji
authors.initials: Y
authors.lastname: Kozuka
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ogawa
authors.forename: Taizo
authors.initials: T
authors.lastname: Shiraishi
authors.forename: Kyoko
authors.initials: K
authors.lastname: Imanaka-Yoshida
mesh_terms: D057134:Antibodies, Neutralizing / Q000494:pharmacology; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology*; D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D005260:Female; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism*; D006801:Humans; D039564:Integrin alphaV / Q000276:immunology; D009361:Neoplasm Invasiveness; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D017154:Stromal Cells / Q000187:drug effects / Q000473:pathology; D013347:Subcellular Fractions / Q000378:metabolism; D019063:Tenascin / Q000494:pharmacology*; D014945:Wound Healing / Q000187:drug effects; D051176:beta Catenin / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D057134:Antibodies, Neutralizing; D015820:Cadherins; D039564:Integrin alphaV; D047428:Protein Kinase Inhibitors; D019063:Tenascin; D051176:beta Catenin; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D019061:src-Family Kinases