Abstract
There is considerable evidence for a relationship between hemostasis and malignancy. Since platelet adhesion to tumor cells has been implicated in the metastatic process and plasma levels of fibrinogen (Fg) and soluble fibrin (sFn) monomer are increased in cancer, we hypothesized that these molecules might enhance tumor-platelet interaction. We therefore studied binding of sFn monomer to tumor cells in a static microplate adhesion assay and determined the effect of pre-treating tumor cells with sFn on tumor cell-induced thrombocytopenia and experimental metastasis. Soluble fibrin (produced by adding thrombin to FXIII- and plasminogen-free Fg in the presence of Gly-Pro-Arg-Pro-amide (GPRP-NH2) significantly increased platelet adherence to tumor cells. This effect was primarily mediated by the integrins alphaIIb beta3 on the platelet and CD 54 (ICAM-1) on the tumor cells. Platelets adhered to untreated A375 cells (28 +/- 8 platelets/tumor cell) and this was not significantly affected by pre-treatment of the tumor cells with fibrinogen or GPRP-NH2. Although thrombin treatment increased adherence, pre-incubation of the tumor cells with sFn resulted in a further increase in platelet binding to tumor cells. In contrast to untreated tumor cells, intravenous injection of sFn-treated A 375 cells reduced the platelet count in anticoagulated mice, supporting the in vitro finding that sFn enhanced tumor cell-platelet adherence. In a more aggressive model of experimental metastasis, treating tumor cells with sFn enhanced lung seeding by 65% compared to untreated cells. Extrapolation of our data to the clinical situation suggests that coagulation activation, and subsequent increase in circulating Fn monomer, may enhance platelet adhesion to circulating tumor cells and thereby facilitate metastatic spread.
MeSH Terms
Animals
Antigens, CD/biosynthesis
Antigens, Human Platelet/biosynthesis
Batroxobin/pharmacology
Blood Platelets/cytology,drug effects
Cell Adhesion/physiology
Cell Communication/physiology
Female
Fibrin/metabolism,pharmacology,physiology
Fibrinolytic Agents/pharmacology
Flow Cytometry
Hemostatics/metabolism,pharmacology
Humans
Lung Neoplasms/secondary
Melanoma, Amelanotic/complications,metabolism,pathology,secondary
Mice
Mice, Nude
Platelet Adhesiveness/drug effects,physiology
Receptors, Thrombin/metabolism
Solubility
Thrombin/metabolism,pharmacology
Thrombocytopenia/drug therapy,etiology
Tumor Cells, Cultured/drug effects
Chemicals
Antigens, CD
Antigens, Human Platelet
Fibrinolytic Agents
Hemostatics
Receptors, Thrombin
human platelet antigen 1b
Fibrin
Batroxobin
Thrombin
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Biggerstaff J P
Research and Clinical Laboratories, Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando 32804, USA.
[email protected]
Seth N
Amirkhosravi A
Amaya M
Fogarty S
Meyer T V
Siddiqui F
Francis J L
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