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Thromb Haemost 2016; 115(05): 1025-1033
DOI: 10.1160/TH15-07-0541
Endothelium and Angiogenesis
Schattauer GmbH

Tissue factor associates with survival and regulates tumour progression in osteosarcoma

Chris Tieken
1   Einthoven Laboratory for Experimental Vascular Medicine, Department of Thrombosis and Hemostasis, Leiden University Medical Centre, Leiden, The Netherlands
,
Michiel C. Verboom
2   Department of Clinical Oncology, Leiden University Medical Centre, Leiden, The Netherlands
,
Wolfram Ruf
3   Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, California, USA
,
Hans Gelderblom
2   Department of Clinical Oncology, Leiden University Medical Centre, Leiden, The Netherlands
,
Judith V. M. G. Bovée
4   Department of Pathology, Leiden University Medical Centre, Leiden, The Netherlands
,
Pieter H. Reitsma
1   Einthoven Laboratory for Experimental Vascular Medicine, Department of Thrombosis and Hemostasis, Leiden University Medical Centre, Leiden, The Netherlands
,
Anne-Marie Cleton-Jansen
4   Department of Pathology, Leiden University Medical Centre, Leiden, The Netherlands
,
Henri H. Versteeg
1   Einthoven Laboratory for Experimental Vascular Medicine, Department of Thrombosis and Hemostasis, Leiden University Medical Centre, Leiden, The Netherlands
› Author Affiliations Financial Support: W. R. is funded by NIH grant HL60742, H. H. V. is funded by NWO grant 91710329.
Further Information

Publication History

Received: 07 July 2015

Accepted after major revision: 02 January 2015

Publication Date:
06 December 2017 (online)

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Summary

Osteosarcoma is the most common primary malignant bone tumour. Patients often develop lung metastasis and have a poor prognosis despite extensive chemotherapy and surgical resections. Tissue Factor is associated with poor clinical outcome in a wide range of cancer types, and promotes angiogenesis and metastasis. The role of Tissue Factor in OS tumourigenesis is unknown. Fifty-three osteosarcoma pre-treatment biopsies and four osteosarcoma cell lines were evaluated for Tissue Factor expression, and a possible association with clinical parameters was investigated. Tissue Factor function was inhibited in an osteosarcoma cell line (143B) by shRNA knockdown or specific antibodies, and pro-tumourigenic gene expression, proliferation, matrigel invasion and transwell migration was examined. 143B cells were implanted in mice in the presence of Tissue Factor-blocking antibodies, and tumour volume, micro-vessel density and metastases in the lung were evaluated. Tissue Factor was highly expressed in 73.6 % of osteosarcoma biopsies, and expression associated significantly with disease-free survival. Tissue Factor was expressed in all four investigated cell lines. Tissue Factor was knocked down in 143B cells, which led to reduced expression of IL-8, CXCL-1, SNAIL and MMP2, but not MMP9. Tissue Factor knockdown or inhibition with antibodies reduced matrigel invasion. Tissue Factor antibodies limited 143B tumour growth in vivo, and resulted in decreased intra-tumoural micro-vessel density. Furthermore, lung metastasis from the primary tumour was significantly reduced. Thus, Tissue Factor expression in osteosarcoma reduces metastasis-free survival in patients, and increases pro-tumourigenic behaviour both in vitro and in vivo.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Osteosarcoma - coagulation - tumour - migration

 

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