Evaluation of procoagulant tissue factor expression in canine hemangiosarcoma cell lines

Abstract

OBJECTIVE To evaluate expression of procoagulant tissue factor (TF) by canine hemangiosarcoma cells in vitro. SAMPLES 4 canine hemangiosarcoma cell lines (SB-HSA [mouse-passaged cutaneous tumor], Emma [primary metastatic brain tumor], and Frog and Dal-1 [primary splenic tumors]) and 1 nonneoplastic canine endothelial cell line (CnAoEC). PROCEDURES TF mRNA and TF antigen expression were evaluated by quantitative real-time PCR assay and flow cytometry, respectively. Thrombin generation was measured in canine plasma and in coagulation factor–replete or specific coagulation factor–deficient human plasma by calibrated automated thrombography. Corn trypsin inhibitor and annexin V were used to examine contributions of contact activation and membrane-bound phosphatidylserine, respectively, to thrombin generation. RESULTS All cell lines expressed TF mRNA and antigen, with significantly greater expression of both products in SB-HSA and Emma cells than in CnAoEC. A greater percentage of SB-HSA cells expressed TF antigen, compared with other hemangiosarcoma cell lines. All hemangiosarcoma cell lines generated significantly more thrombin than did CnAoEC in canine or factor-replete human plasma. Thrombin generation induced by SB-HSA cells was significantly lower in factor VII-deficient plasma than in factor-replete plasma and was abolished in factor X–deficient plasma; residual thrombin generation in FVII-deficient plasma was abolished by incubation of cells with annexin V. Thrombin generation by SB-HSA cells was unaffected by the addition of corn trypsin inhibitor. CONCLUSIONS AND CLINICAL RELEVANCE Hemangiosarcoma cell lines expressed procoagulant TF in vitro. Further research is needed to determine whether TF can be used as a biomarker for hemostatic dysfunction in dogs with hemangiosarcoma.

The prevalence of intra‐tumoral and distant thrombi, as well as tumour‐cell emboli in canine neoplasia

“…52,53 Other solid tumours that have also been associated with haemostatic dysfunction includes carcinomas and lymphomas. 53,54 The blood pooling in a disorganized vasculature, procoagulant factor expression on the tumour vessel wall, altered blood flow within the neoplasm and a pro-inflammatory state of the affected endothelium within the neoplasm, [55][56][57][58][59] may result in upregulation of the haemostasis pathways and the subsequent development of a hypercoagulable state, likely resulting in intra-tumoral thrombi. 20,27,36 Intra-tumoral thrombosis can result in compromised blood flow, ischemia, and ultimately necrosis.…”

A Novel Microchip Flow Chamber (Total Thrombus Analysis System) to Assess Canine Hemostasis

“…Similarly, it is not known if all of the dogs with the bleeding disorders studied are truly representative of their condition. This is particularly the case with dogs that have acquired bleeding disorders such as those dogs with hemangiosarcoma since the bleeding disorders associated with cancer can be varied and complex (60)(61)(62). Two of the healthy control dogs in the present study each had one abnormal coagulation value (increased Ddimer or increased VWF:Ag).…”

In vitro effects of the chemotherapy agent water‐soluble micellar paclitaxel (Paccal Vet) on canine hemangiosarcoma cell lines

“…Two canine HSA cell lines, DEN‐HSA (provided by Dr Ilene Kurzman, University of Wisconsin, Madison, Wisconsin) and Frog (donated from Dr Jaime Modiano, University of Minnesota, Saint Paul, Minnesota) were used. Both HSA cell lines have been characterized and reported before . Each cell line was maintained in 75 cm 2 plastic flasks in 10 mL of minimum essential media (MEM) (Corning cellgro, Manassas, Virginia) and incubated at 37°C under 5% CO 2 .…”