Temozolomide and Lomustine Induce Tissue Factor Expression and Procoagulant Activity in Glioblastoma Cells In Vitro

Cancers, vol. 15, pp. 2347

Abstract

Glioblastoma (GBM) patients have one of the highest risks of venous thromboembolism (VTE), which is even further increased upon treatment with chemotherapy. Tissue factor (TF) is the initiator of the extrinsic coagulation pathway and expressed by GBM cells. In this study, we aimed to examine the effect of routinely used chemotherapeutic agents Temozolomide (TMZ) and Lomustine (LOM) on TF procoagulant activity and expression in GBM cells in vitro. Three human GBM cell lines (U-251, U-87, U-118) were exposed to 100 µM TMZ or 30 µM LOM for 72 h. TF procoagulant activity was assessed via an FXa generation assay and TF gene and protein expression through qPCR and Western blotting. The externalization of phosphatidylserine (PS) was studied using Annexin V flow cytometry. Treatment with TMZ and LOM resulted in increased procoagulant activity in all cell lines. Furthermore, both agents induced procoagulant activity in the supernatant and tumor-cell-secreted extracellular vesicles. In line, TF gene and protein expression were increased upon TMZ and LOM treatment. Additionally, PS externalization and induction of inflammatory-associated genes were observed. Overall, the chemotherapeutic modalities TMZ and LOM induced procoagulant activity and increased TF gene and protein expression in all GBM cell lines tested, which may contribute to the increased VTE risk observed in GBM patients undergoing chemotherapy.

Authors 10

1. Maaike Y. Kapteijn Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands

2. Shanna Zwaan Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands

3. Esther ter Linden Aachen Department of Cell and Chemical Biology Oncode Institute

   Leiden University Medical Center · Oncode Institute

   Affiliation as printed

   Department of Cell and Chemical Biology, ONCODE Institute, Leiden University Medical Center, 2333 ZC Leiden, The Netherlands

4. El Houari Laghmani Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands

5. Rob F. P. van den Akker Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands

6. Araci M. R. Rondon Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands

7. Sabina Y. van der Zanden Aachen Department of Cell and Chemical Biology Oncode Institute

   Leiden University Medical Center · Oncode Institute

   Affiliation as printed

   Department of Cell and Chemical Biology, ONCODE Institute, Leiden University Medical Center, 2333 ZC Leiden, The Netherlands

8. Jacques Neefjes Aachen Department of Cell and Chemical Biology Oncode Institute

   Leiden University Medical Center · Oncode Institute

   Affiliation as printed

   Department of Cell and Chemical Biology, ONCODE Institute, Leiden University Medical Center, 2333 ZC Leiden, The Netherlands

9. Henri H. Versteeg Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands

10. Jeroen T. Buijs corresponding Aachen Einthoven Laboratory for Vascular and Regenerative Medicine Division of Thrombosis and Hemostasis Department of Internal Medicine

    Leiden University · Leiden University Medical Center

    Affiliation as printed

    Einthoven Laboratory for Vascular and Regenerative Medicine, Division of Thrombosis & Hemostasis, Department of Internal Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands