Platelet adhesion in breast cancer: development and application of a novel assay.
The increased risk of thromboembolism in cancer may be related to a prothrombotic or hypercoagulable state, with abnormalities of haemostasis and platelet activation. To further investigate the role of platelets in this disease, we developed and applied a new assay to detect and quantify platelet adhesion to the well-defined subendothelial substrate, fibrinogen. Platelet-rich plasma was obtained from 31 females with breast cancer (13 metastatic, 18 benign), and 30 healthy female controls, re-suspended to 2 x 10(8) cells/ml and 100 microl and incubated for 1 h in microtitre plates pre-coated with fibrinogen (5 mg/ml). The supernatant was carefully aspirated, lysed with Triton X-100 and stored at -70 degrees C as supernatant-platelet lysate. The microtitre wells were carefully washed with saline, bound platelets lysed with Triton, and the lysate stored at -70 degrees C as bound-platelet lysate. P-selectin was determined in supernatant-platelet lysate and bound-platelet lysate for each patient by enzyme-linked immunosorbent assay. Interpreting differences in P-selectin in different lysates as reflective of adhesion, patients with cancer had increased platelet adhesion (absolute and percentage, both P < 0.001) compared with healthy controls. There was also more adhesion (P < 0.001) in metastatic disease compared with non-metastatic disease. Patients with breast carcinomas, and, in particular, those with metastatic disease, have a higher degree of platelet adhesion, which may by quantified by a novel method based on cell lysis. This increase in platelet adhesiveness may be related to an increased risk of thromboembolism in these patients.
Caine
Graham J
GJ
Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, UK.
Nadar
Sunil K
SK
Lip
Gregory Y H
GY
Stonelake
Paul S
PS
Blann
Andrew D
AD
D001792:Blood Platelets / Q000187:drug effects; D001941:Breast Diseases / Q000097:blood; D001943:Breast Neoplasms / Q000097:blood*; D002277:Carcinoma / Q000097:blood*; D015331:Cohort Studies; D003902:Detergents / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005340:Fibrinogen; D006801:Humans; D008875:Middle Aged; D009362:Neoplasm Metastasis; D017830:Octoxynol / Q000494:pharmacology; D019007:P-Selectin / Q000032:analysis; D010865:Pilot Projects; D010949:Plasma; D010973:Platelet Adhesiveness; D010979:Platelet Function Tests
D003902:Detergents; D019007:P-Selectin; D005340:Fibrinogen; D017830:Octoxynol