Clin Res Cardiol 101, Suppl 1, April 2012

P368 - Characterization of the pro-angiogenic properties of alternatively spliced human Tissue Factor in human cells
A. Eisenreich1, H.-P. Schultheiss1, U. Rauch-Kröhnert1
1Med. Klinik II, Klinik für Kardiologie und Pulmologie, Charité - Universitätsmedizin Berlin, Campus Benj. Franklin, Berlin;
Background: Tissue factor (TF) is a transmembrane glycoprotein, the primary initiator of blood coagulation and was suggested to play a role in angiogenesis. Two naturally occurring TF isoforms are expressed in human cells, soluble alternatively spliced (as)TF and “full length” (fl)TF. Recent reports indicate the alternatively spliced TF isoform to be involved in organ growth and angiogenesis in a mouse model and to be a potential pro-angiogenic protein in human cells. How asTF influences angiogenesis in human tissues, such as the heart and lung is still unknown. It is possible that the soluble TF isoform may act via interactions with receptors, such as integrins or other factors which are involved in pro-angiogenic processes, such as the chemokines MCP-1 and Cyr61 or VEGF165. Here, we investigated the impact of stable asTF overexpression in the human cell line A549 on the pro-angiogenic properties and the function of these cells, such as cell migration and proliferation.
Methods: A549 cells were stable transfected with an asTF-overexpression plasmid. AsTF overexpression was verified by TF isoform-specific Real-Time PCR. The expression of several pro-angiogenic factors, such as VEGF165, Cyr61 or MCP-1 was assed by semi-quantitative RT-PCR and Western blotting. The cell migration and proliferation was determined via appropriate assays in vitro.
Results: A549 cells expressed the pro-angiogenic factors: VEGF165, Cyr61 and MCP-1. On mRNA level the expression of VEGF165 was 1.5-fold increased in asTF-overexpressing A459 cells compared to cells transfected with a control plasmid (p<0.05). Cyr61 was approximately 2-fold increased (p<0.001) and MCP-1 was 8-fold increased in asTF-overexpressing cells compared to the control cells (p<0.05). The protein expression of all the pro-angiogenic factors was also induced in asTF-overexpressing cells compared to controls. Also the cellular function was modified by asTF overexpression. The cell proliferation of A549 cells was significantly increased in asTF-overexpressing cells compared to controls (p<0.05). Furthermore, the increase in migration of monocytes treated with the supernatant of asTF-overexpressing A549 cells was comparable to controls treated with the pro-migratory chemo attractant MCP-1.
Conclusion: These results indicated asTF to increase the expression of different pro-angiogenic factors and to induce the cell proliferation of human cells as well as the migration of human monocytes. Thus, asTF is a potent pro-angiogenic agent, contributing to angiogenesis in human organs.
Clin Res Cardiol 101, Suppl 1, April 2012
Zitierung mit Vortrags- oder Posternummer s.o.
DOI 10.1007/s00392-012-1100-6