Tissue Factor Potentiates The Adherence Of Breast Cancer Cells To Human Umbilical Vein Endothelial Cells

ObjectiveThe aim of this study is to investigate the potential roles of tissue factor in the cell adhesion between breast cancer cells and HUVECs examined by static adhesion assay and parallel plate flow assay,this study will provide a molecular basis for the future tumor metastasis targeted therapy.Methods1.The expression of TF in three breast cancer cell lines(MDA-MB-231,HS-578 T,MCF-7)and Human umbilical vein endothelial cells,the expression of integrinβ1 in MDA-MB-231 and HS-578 T,the xpression of CD44,β1 Integrin and ICAM-1 in HS-578 T Cells pretreated with TF recombinant protein were detected by Western-Blotting analysis.2.The expression of TF in breast cancer cell lines(MDA-MB-231,HS-578 T,MCF-7)was detected by flow cytometry analysis.3.The expression and location of TF in MDA-MB-231 breast cancer cells were examined by immunohistochemical staining.4.Static Adhesion assay and Parallel plate flow assay were used to study three breast cancer cells lines adhere to human umbilical vein endothelial cells;Endothelial cell adhesion assay was used to detect endothelial cell adhesion to TF recombinant protein.5.MTT method was used to detect the viability of cells pretreated with TF antibody integrinβ1 antibody and recombinant TF protein.Results1.The expression of TF in MDA-MB-231 cells was significantly higher than that in HS-578 T and MCF-7 cells.2.The adhesion between MDA-MB-231 cells and HUVEC was higher than HS-578 T and MCF-7 cells(P<0.05),the adhesion between MDA-MB-231 cells blocked with TF antibody and HUVEC decreased compared with control(P<0.05).3.The adhesion of HS-578 T cells pretreated with recombinant tissue factor protein to HUVEC increased compared with unpretreated HS-578 T cells(P<0.05).4.Human umbilical vein endothelial cells expressed low levels of TF and high levels of β1integrin,Human umbilical vein endothelial cells adhesion to tissue factor recombinant protein increased compared with control(P<0.05).When blocked with β1 integrin antibody,the adhesion decreased(P<0.05),the adhesion of MDA-MB-231 blocked with TF antibody to HUVEC pretreated with β1 integrin antibody was decreased(P<0.05).5.The expression of CD44 in MDA-MB-231 cells was higher than that in HS-578 T cells(P<0.05),the expression of CD44 was increased in HL-578 T cells treated with recombinant tissue factor protein(P<0.05).Conclusion1.The adhesion of MDA-MB-231 expressing high levels of tissue factor to HUVEC is increased,and the cell adhesion between HS-578 T and HUVEC is increased pretreated with recombinant tissue factor protein,It suggest that TF may mediate the adhesion of breast cancer cell adhesion to HUVEC.2.It suggest that the tissue factor on the surface of MDA-MB-231 cells may interact with β1integrin on HUVEC mediated the cell adhesion.3.The expression of CD44 adhesion moleculor is higher in MDA-MB-231 than that in HS-578 T cells,the expression of CD44 in HS-578 T increased after pretreated with recombinant protein TF,and consistent with the high adhesion ability of MDA-MB-231 cells and the enhanced adhesion ability of HS-578 T cells treated with recombinant protein TF,it suggest that TF could upregulate the expression of CD44 mediating the adhesion between breast cancer cells and HUVEC.