Platelet adhesion in breast cancer: Development and application of a novel assay

Graham J. Caine, Sunil K. Nadar, Gregory Y H Lip, Paul S. Stonelake, Andrew D. Blann

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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 × 10 8 cells/ml and 100 μl 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°C as supernatant-platelet lysate. The microtitre wells were carefully washed with saline, bound platelets lysed with Triton, and the lysate stored at -70°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.

Original languageEnglish
Pages (from-to)513-518
Number of pages6
JournalBlood Coagulation and Fibrinolysis
Volume15
Issue number6
DOIs
Publication statusPublished - Sep 2004

Fingerprint

Blood Platelets
Breast Neoplasms
P-Selectin
Thromboembolism
Fibrinogen
Platelet Adhesiveness
Platelet-Rich Plasma
Platelet Activation
Octoxynol
Hemostasis
Neoplasms
Enzyme-Linked Immunosorbent Assay

Keywords

  • Adhesion
  • Enzyme-linked immunosorbent assay
  • Fibrinogen
  • P-selectin
  • Platelets

ASJC Scopus subject areas

  • Hematology

Cite this

Platelet adhesion in breast cancer : Development and application of a novel assay. / Caine, Graham J.; Nadar, Sunil K.; Lip, Gregory Y H; Stonelake, Paul S.; Blann, Andrew D.

In: Blood Coagulation and Fibrinolysis, Vol. 15, No. 6, 09.2004, p. 513-518.

Research output: Contribution to journalArticle

Caine, Graham J. ; Nadar, Sunil K. ; Lip, Gregory Y H ; Stonelake, Paul S. ; Blann, Andrew D. / Platelet adhesion in breast cancer : Development and application of a novel assay. In: Blood Coagulation and Fibrinolysis. 2004 ; Vol. 15, No. 6. pp. 513-518.
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