TY - JOUR TI - The Lectin-Like Domain of Thrombomodulin Inhibits β1 Integrin-Dependent Binding of Human Breast Cancer-Derived Cell Lines to Fibronectin AU - Kawamoto, Eiji AU - Nago, Nodoka AU - Okamoto, Takayuki AU - Gaowa, Arong AU - Masui-Ito, Asami AU - Akama, Yuichi AU - Darkwah, Samuel AU - Appiah, Michael Gyasi AU - Myint, Phyoe Kyawe AU - Obeng, Gideon AU - Ito, Atsushi AU - Caidengbate, Siqingaowa AU - Esumi, Ryo AU - Yamaguchi, Takanori AU - Park, Eun Jeong AU - Imai, Hiroshi AU - Shimaoka, Motomu T2 - Biomedicines AB - Thrombomodulin is a molecule with anti-coagulant and anti-inflammatory properties. Recently, thrombomodulin was reported to be able to bind extracellular matrix proteins, such as fibronectin and collagen; however, whether thrombomodulin regulates the binding of human breast cancer-derived cell lines to the extracellular matrix remains unknown. To investigate this, we created an extracellular domain of thrombomodulin, TMD123-Fc, or domain deletion TM-Fc proteins (TM domain 12-Fc, TM domain 23-Fc) and examined their bindings to fibronectin in vitro by ELISA. The lectin-like domain of thrombomodulin was found to be essential for the binding of the extracellular domain of thrombomodulin to fibronectin. Using a V-well cell adhesion assay or flow cytometry analysis with fluorescent beads, we found that both TMD123-Fc and TMD12-Fc inhibited the binding between β1 integrin of human breast cancer-derived cell lines and fibronectin. Furthermore, TMD123-Fc and TMD12-Fc inhibited the binding of activated integrins to fibronectin under shear stress in the presence of Ca2+ and Mg2+ but not under strong integrin-activation conditions in the presence of Mg2+ without Ca2+. This suggests that thrombomodulin Fc fusion protein administered exogenously at a relatively early stage of inflammation may be applied to the development of new therapies that inhibit the binding of β1 integrin of breast cancer cell lines to fibronectin. DA - 2021/02// PY - 2021 DO - 10.3390/biomedicines9020162 DP - www.mdpi.com VL - 9 IS - 2 SP - 162 LA - en UR - https://www.mdpi.com/2227-9059/9/2/162 Y2 - 2021/02/10/23:42:53 KW - V-well assay KW - breast cancer cell KW - cell adhesion KW - integrin KW - thrombomodulin ER -