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Published on: August 3, 2018
Adenovirus-mediated expression of tissue factor pathway inhibitor-2 inhibits endothelial cell migration and
Lacramioara Ivanciu1, Robert D Gerard, Haiwang Tang
1Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK 73104, USA.
Objective:
Extracellular matrix (ECM) remodeling during angiogenesis is accomplished through plasmin-dependent pericellular proteolysis and through the action of matrix metalloproteinases (MMPs). Because tissue factor pathway inhibitor-2 (TFPI-2), a Kunitz-type protease inhibitor with prominent ECM localization, inhibits plasmin and MMPs activity, we investigated the role of TFPI-2 in endothelial cell (EC) migration and angiogenesis.
Methods And Results:
Real-time polymerase chain reaction and immunostaining showed that the expression of TFPI-2 mRNA and protein was upregulated in migrating ECs. The effect of TFPI-2 on angiogenesis was studied in mouse models of Matrigel and polyvinylalcohol sponge implants by overexpressing TFPI-2 through infection with a replication-deficient adenovirus (AdTFPI-2). Using (immuno)fluorescence and confocal microscopy we observed that TFPI-2 reduced neovascularization and promoted ECM deposition. Lateral cell migration and capillary tube formation in vitro also were impaired by TFPI-2, a process reversed by anti-TFPI-2 antibodies. Increased apoptosis occurred both in AdTFPI-2-treated ECs and in the mouse implants. Zymography and assays in the absence of plasminogen confirmed plasmin inhibition as a main mechanism through which TFPI-2 inhibits EC migration.
Conclusions:
Our data suggest that TFPI-2 may be an important regulator of aberrant angiogenesis associated with tumor growth/metastasis, cardiovascular diseases, chronic inflammation, or diabetes.
Insights
Tissue factor pathway inhibitor-2 (TFPI-2) impairs endothelial cell migration and angiogenesis by inhibiting plasmin. This suggests TFPI-2 plays a role in diseases involving abnormal blood vessel growth.
Area of Science:
- Molecular biology
- Cell biology
- Biochemistry
Background:
- Extracellular matrix (ECM) remodeling is crucial for angiogenesis.
- Plasmin and matrix metalloproteinases (MMPs) are key enzymes in ECM remodeling.
- Tissue factor pathway inhibitor-2 (TFPI-2) inhibits plasmin and MMPs and is found in the ECM.
Purpose of the Study:
- To investigate the role of TFPI-2 in endothelial cell (EC) migration and angiogenesis.
- To understand the mechanisms by which TFPI-2 affects these processes.
Main Methods:
- Real-time PCR and immunostaining to assess TFPI-2 expression in migrating ECs.
- Mouse models (Matrigel and polyvinylalcohol sponge implants) with TFPI-2 overexpression via adenovirus.
- In vitro assays for EC migration and capillary tube formation.
- Zymography and assays without plasminogen to confirm plasmin inhibition.
Main Results:
- TFPI-2 expression was upregulated in migrating ECs.
- Overexpression of TFPI-2 reduced neovascularization, promoted ECM deposition, and impaired EC migration and capillary tube formation in vitro.
- TFPI-2 inhibition of plasmin was confirmed as a primary mechanism.
- Increased apoptosis was observed in TFPI-2-treated ECs and implants.
Conclusions:
- TFPI-2 is an important regulator of angiogenesis.
- TFPI-2's role in inhibiting plasmin contributes to its anti-angiogenic effects.
- TFPI-2 may be a significant factor in aberrant angiogenesis related to tumors, cardiovascular diseases, inflammation, and diabetes.
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