title: Hypoxia regulates the expression of tissue factor pathway signaling elements in a rat glioma model.
abstract: Hypoxia and necrosis are fundamental features of glioma, and their emergence is critical for the rapid biological progression of this fatal tumor. The presence of vaso-occlusive thrombus is higher in grade IV tumors [glioblastoma multiforme (GBM)] compared with lower grade tumors, suggesting that the procoagulant properties of the tumor contribute to its aggressive behavior, as well as the establishment of tumor hypoxia and necrosis. Tissue factor (TF), the primary cellular initiator of coagulation, is overexpressed in GBMs and likely favors a thrombotic microenvironment. Phosphatase and tensin homolog (PTEN) loss and hypoxia are two common alterations observed in glioma that may be responsible for TF upregulation. In the present study, ST1 and P7 rat glioma lines, with different levels of aggressiveness, were comparatively analyzed with the aim of identifying differences in procoagulant mechanisms. The results indicated that P7 cells display potent procoagulant activity compared with ST1 cells. Flow cytometric analysis showed less pronounced levels of TF in ST1 cells compared with P7 cells. Notably, P7 cells supported factor X (FX) activation via factor VIIa, whereas no significant FXa generation was observed in ST1 cells. Furthermore, the exposure of phosphatidylserine on the surface of P7 and ST1 cells was investigated. The results supported the assembly of prothrombinase complexes, accounting for the production of thrombin. Furthermore, reverse transcription-quantitative polymerase chain reaction showed that CoCl2(known to induce a hypoxic-like stress) led to an upregulation of TF levels in P7 and ST1 cells. Therefore, increased TF expression in P7 cells was accompanied by increased TF procoagulant activity. In addition, hypoxia increased the shedding of procoagulant TF-bearing microvesicles in both cell lines. Finally, hypoxic stress induced by treatment with CoCl2upregulated the expression of the PAR1 receptor in both P7 and ST1 cells. In addition to PAR1, P7, but not ST1 cells, expressed higher levels of PAR2 under hypoxic stress. Thus, modulating these molecular interactions may provide additional insights for the development of more efficient therapeutic strategies against aggressive glioma.
authors.affiliation: Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.
authors.forename: Robson Q
authors.initials: RQ
authors.lastname: Monteiro
authors.affiliation: Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil; Bone Marrow Transplantation Center, National Institute of Cancer, Rio de Janeiro, RJ 20230-130, Brazil.
authors.forename: Luize G
authors.initials: LG
authors.lastname: Lima
authors.affiliation: Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.
authors.forename: Nathália P
authors.initials: NP
authors.lastname: Gonçalves
authors.affiliation: Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.
authors.forename: Mayara R Arruda
authors.initials: MR
authors.lastname: DE Souza
authors.affiliation: Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Leal
authors.affiliation: Cell and Molecular Therapy Center (NUCEL-NETCEM), Internal Medicine Department, School of Medicine, University of São Paulo, São Paulo, SP 05360-120, Brazil.
authors.forename: Marcos A Almeida
authors.initials: MA
authors.lastname: Demasi
authors.affiliation: Cell and Molecular Therapy Center (NUCEL-NETCEM), Internal Medicine Department, School of Medicine, University of São Paulo, São Paulo, SP 05360-120, Brazil.
authors.forename: Mari C
authors.initials: MC
authors.lastname: Sogayar
authors.affiliation: Institute of Medical Biochemistry Leopoldo de Meis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil.
authors.forename: Tatiana C
authors.initials: TC
authors.lastname: Carneiro-Lobo
keywords: blood coagulation; glioma; hypoxia; microvesicles; protease-activated receptor; tissue factor; vascular endothelial growth factor