Daunorubicin induces procoagulant activity of cultured endothelial cells through phosphatidylserine exposure and microparticles release

Yueyue Fu; Jin Zhou; Huibo Li; Fenglin Cao; Yanhua Su; Shengjin Fan; Yinghua Li; Shuye Wang; Limin Li; Gary E. Gilbert; Jialan Shi

doi:10.1160/TH10-02-0102

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2010; 104(06): 1235-1241
DOI: 10.1160/TH10-02-0102

Cellular Proteolysis and Oncology

Schattauer GmbH

Daunorubicin induces procoagulant activity of cultured endothelial cells through phosphatidylserine exposure and microparticles release

Yueyue Fu^*

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Jin Zhou^*

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Huibo Li

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Fenglin Cao

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Yanhua Su

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Shengjin Fan

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Yinghua Li

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Shuye Wang

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Limin Li

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

,

Gary E. Gilbert

²Medicine Departments of VA Boston Healthcare System, Brigham and Women’s Hospital, and Harvard Medical School, Boston, Massachusetts, USA

,

Jialan Shi

¹Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China

²Medicine Departments of VA Boston Healthcare System, Brigham and Women’s Hospital, and Harvard Medical School, Boston, Massachusetts, USA

› Author Affiliations

Abstract

Summary

Administration of various chemotherapeutic agents is associated with an increased risk of thrombotic events. Although vascular endothelium plays a predominant role in blood coagulation and fibrinolysis, the effect of cytotoxic drugs on the procoagulant activity (PCA) of endothelial cells has not been well evaluated. Our study aims to investigate the possibility that daunorubicin, a chemotherapeutic agent, exerts prothrombotic effect on endothelial cells. We tested the impact of daunorubicin on phosphatidylserine (PS) exposure, endothelial microparticles (EMPs) release and consequent PCA. Human umbilical vein endothelial cells (HUVECs) were treated with daunorubicin (0.1, 0.2, 0.5, 1, 2 μM) for 24 hours. PCA of HUVECs was measured using clotting time and purified coagulation complex assays. Counts and PCA of EMPs were evaluated by flow cytometry and clotting time assay, respectively. Lactadherin was used as a novel probe for detection of PS exposure and EMPs release. We found that daunorubicin dose-dependently increased the PS exposure and consequent PCA of HUVECs. Moreover, daunorubicin treatment also enhanced the release of EMPs which were highly procoagulant. This increment was especially significant at 0.2 μM of daunorubicin or more. Blockade of PS with lactadherin inhibited over 90% of HUVECs and EMPs PCA. However, anti-TF antibody had no significant inhibition effect. Our results demonstrate that daunorubicin treatment enhanced PCA of HUVECs through PS exposure and shedding of procoagulant EMPs. Lactadherin acts as an efficient anticoagulant in this process.

Keywords

Daunorubicin - endothelial cells - phosphatidylserine - microparticles - lactadherin

Full Text

References

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