TY - JOUR
T1 - Endothelial integrin α3β1 stabilizes carbohydrate-mediated tumor/endothelial cell adhesion and induces macromolecular signaling complex formation at the endothelial cell membrane
AU - Glinskii, Olga V.
AU - Li, Feng
AU - Wilson, Landon S.
AU - Barnes, Stephen
AU - Rittenhouse-Olson, Kate
AU - Barchi, Joseph J.
AU - Pienta, Kenneth J.
AU - Glinsky, Vladislav V.
PY - 2014
Y1 - 2014
N2 - Blood borne metastatic tumor cell adhesion to endothelial cells constitutes a critical rate-limiting step in hematogenous cancer metastasis. Interactions between cancer associated carbohydrate Thomsen-Friedenreich antigen (TF-Ag) and endothelium-expressed galectin-3 (Gal-3) have been identified as the leading molecular mechanism initiating tumor/endothelial cell adhesion in several types of cancer. However, it is unknown how these rather weak and transient carbohydrate/ lectin mediated interactions are stabilized. Here, using Western blot and LC tandem mass spectrometry analyses of pull-downs utilizing TF-Ag loaded gold nanoparticles, we identified Gal-3, endothelial integrin a3ß1, Src kinase, as well as 5 additional molecules mapping onto focal adhesion pathway as parts of the macromolecular complexes formed at the endothelial cell membranes downstream of TF-Ag/Gal- 3 interactions. In a modified parallel flow chamber assay, inhibiting a3ß1 integrin greatly reduced the strength of tumor/endothelial cell interactions without affecting the initial cancer cell adhesion. Further, the macromolecular complex induced by TF-Ag/Gal-3/a3ß1 interactions activates Src kinase, p38, and ERK1/2, pathways in endothelial cells in a time- and a3ß1-dependent manner. We conclude that, following the initial metastatic cell attachment to endothelial cells mediated by TF-Ag/Gal-3 interactions, endothelial integrin a3ß1 stabilizes tumor/endothelial cell adhesion and induces the formation of macromolecular signaling complex activating several major signaling pathways in endothelial cells.
AB - Blood borne metastatic tumor cell adhesion to endothelial cells constitutes a critical rate-limiting step in hematogenous cancer metastasis. Interactions between cancer associated carbohydrate Thomsen-Friedenreich antigen (TF-Ag) and endothelium-expressed galectin-3 (Gal-3) have been identified as the leading molecular mechanism initiating tumor/endothelial cell adhesion in several types of cancer. However, it is unknown how these rather weak and transient carbohydrate/ lectin mediated interactions are stabilized. Here, using Western blot and LC tandem mass spectrometry analyses of pull-downs utilizing TF-Ag loaded gold nanoparticles, we identified Gal-3, endothelial integrin a3ß1, Src kinase, as well as 5 additional molecules mapping onto focal adhesion pathway as parts of the macromolecular complexes formed at the endothelial cell membranes downstream of TF-Ag/Gal- 3 interactions. In a modified parallel flow chamber assay, inhibiting a3ß1 integrin greatly reduced the strength of tumor/endothelial cell interactions without affecting the initial cancer cell adhesion. Further, the macromolecular complex induced by TF-Ag/Gal-3/a3ß1 interactions activates Src kinase, p38, and ERK1/2, pathways in endothelial cells in a time- and a3ß1-dependent manner. We conclude that, following the initial metastatic cell attachment to endothelial cells mediated by TF-Ag/Gal-3 interactions, endothelial integrin a3ß1 stabilizes tumor/endothelial cell adhesion and induces the formation of macromolecular signaling complex activating several major signaling pathways in endothelial cells.
KW - Adhesion
KW - Galectin
KW - Integrin
KW - Thomsen-friedenreich antigen
KW - Tumor metastasis
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U2 - 10.18632/oncotarget.1837
DO - 10.18632/oncotarget.1837
M3 - Article
C2 - 24675526
AN - SCOPUS:84898621028
SN - 1949-2553
VL - 5
SP - 1382
EP - 1389
JO - Oncotarget
JF - Oncotarget
IS - 5
ER -