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.
Haemostasis, Thrombosis and Vascular Biology Unit, University Department of Medicine, City Hospital, Birmingham, UK.
Graham J
GJ
Caine
Sunil K
SK
Nadar
Gregory Y H
GY
Lip
Paul S
PS
Stonelake
Andrew D
AD
Blann
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