The role of tissue factor -β1- Integrin complex formation on Src activity during the apoptosis mechanism in endothelial cells

Abstract EN

Tissue factor (TF) is expressed by cells and tissues, specifically endothelial cells following activation.

In addition, an overexpression of TF in the endothelium contributes to a variety of chronic pathological conditions including thrombosis in cancer, metastasis, angiogenesis and cardiovascular disease.

The aim of this study was to investigate the mechanisms of apoptosis in endothelial cells through the TF–β1 integrin complex formation.

Throughout the study, human dermal blood endothelial cells (HDBECs) were transfected with plasmid to express wild-TF or alternatively, transfected to express mutant-TF.

Other cells incubated with green fluorescent protein (t-GFP) were used as a plasmid control.

Inhibition of the β1-integrin was enhanced by treatment with 10 μg/ml of the β1-integrin inhibitor in three samples to test the effect of the β1-integrin inhibitor on wild-TF, mutant-TFAla253 and t-GFP.

Moreover, samples were run on sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) followed by western blotting for Src antibody detection, using phospho-Src and total Src.

The ratio of Src activity for the samples was obtained by dividing phospho-Src by total Src.

The results revealed a significant effect of the β1 integrin inhibition on Src phosphorylation.

The samples that were treated with the β1 integrin inhibitor showed lower Src activity ratios.

Specifically, the cell sample that was transfected to express mutant-TFAla253 had the lowest Src activity among the HDBEC samples treated with the β1-integrin inhibitor.

In contrast, the Src activity ratios were higher in cell samples that were not treated with the β1-integrin inhibitor.

Therefore, both the β1-integrin inhibitor and the mutant-TFAla253 were major factors inducing apoptosis.

The results suggested that the β1-integrin affects TF mediated apoptosis.

The results also suggested that Src is reduced during apoptosis through the β1-integrin inhibition.