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Thromb Haemost 2018; 118(03): 480-489
DOI: 10.1055/s-0038-1629901
Coagulation and Fibrinolysis
Schattauer GmbH Stuttgart

Effects of Low- and High-Dose Chemotherapy Agents on Thrombogenic Properties of Extracellular Vesicles Derived from Breast Cancer Cell Lines

Anat Aharon
,
Anni Rebibo Sabbah
,
Liron Issman
,
Hila Berkovich
,
Reem Copty
,
Yeshayahu Talmon
,
Benjamin Brenner
 Grants or Other Financial Support This study was supported by the Israel Science Foundation (ISF), grant no. 1413/21, and Rappaport Family Institute Grant 2012–2014.
Weitere Informationen

Publikationsverlauf

11. September 2017

02. Januar 2018

Publikationsdatum:
28. Februar 2018 (online)

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Abstract

Background The involvement of extracellular vesicles (EVs) in cancer-associated thrombosis (CT) is unclear. This study aimed to explore the properties of EVs derived from breast cancer (BC) cells following exposure to high- or low-dose chemotherapeutic agents and evaluate thrombogenic effects of these EVs on endothelial cells (ECs).

Methods EVs were isolated from BC cell lines (non-metastatic MCF7, high-metastatic MDA-MB-231), pre-exposed to serum-free medium (control), with or without increasing doses of doxorubicin or paclitaxel. EV structure and size were studied using electron microscopy and Nano-sight. Antigen levels were measured by fluorescence-activated cell sorting (FACS). EV effects on EC thrombogenicity were assessed using FACS, factor Xa chromogenic assay and RT-PCR.

Results Serum-free medium BC cell resulted in EV shedding that additionally increased when MDA-MB-231 cells were exposed to high doses of both agents. Tissue factor (TF) levels were similarly low (9–13%) in all EVs compared with the high expression on their parental MDA-MB-231 cells (76–83%). EVs derived from MDA-MB-231 cells stimulated with high-dose doxorubicin demonstrated significantly (fivefold; p < 0.001) elevated levels of negatively charged phospholipids, a 97% decrease in TF pathway inhibitor (TFPI) levels and a sixfold increase (p < 0.001) in procoagulant activity. These EVs also enhanced EC thrombogenicity. Effects of EVs originating from MCF7 cells were less pronounced.

Conclusion These findings suggest that thrombogenic properties of BC-derived EVs may depend on the type and dose of the applied chemotherapy agent and may also be affected by the cell metastatic nature.

Keywords

extracellular vesicles - breast cancer cells - endothelial cells - high-dose chemotherapy - thrombogenicity
 

  • References

  • 1 Furie B, Furie BC. Mechanisms of thrombus formation. N Engl J Med 2008; 359 (09) 938-949
    CrossrefPubMedGoogle Scholar
  • 2 Zwicker JI, Liebman HA, Neuberg D. , et al. Tumor-derived tissue factor-bearing microparticles are associated with venous thromboembolic events in malignancy. Clin Cancer Res 2009; 15 (22) 6830-6840
    CrossrefPubMedGoogle Scholar
  • 3 DeSantis CE, Lin CC, Mariotto AB. , et al. Cancer treatment and survivorship statistics, 2014. CA Cancer J Clin 2014; 64 (04) 252-271
    CrossrefPubMedGoogle Scholar
  • 4 Hsu JL, Hung MC. The role of HER2, EGFR, and other receptor tyrosine kinases in breast cancer. Cancer Metastasis Rev 2016; 35 (04) 575-588
    CrossrefPubMedGoogle Scholar
  • 5 Yu DD, Wu Y, Shen HY. , et al. Exosomes in development, metastasis and drug resistance of breast cancer. Cancer Sci 2015; 106 (08) 959-964
    CrossrefPubMedGoogle Scholar
  • 6 Paulus JK, Rosenberg AS. Breast cancer and thrombosis: timing matters. Blood 2016; 127 (07) 793-794
    CrossrefPubMedGoogle Scholar
  • 7 Walker AJ, West J, Card TR, Crooks C, Kirwan CC, Grainge MJ. When are breast cancer patients at highest risk of venous thromboembolism? A cohort study using English health care data. Blood 2016; 127 (07) 849-857 , quiz 953
    CrossrefPubMedGoogle Scholar
  • 8 Cole M, Bromberg M. Tissue factor as a novel target for treatment of breast cancer. Oncologist 2013; 18 (01) 14-18
    CrossrefPubMedGoogle Scholar
  • 9 Aharon A, Brenner B. Microparticles, thrombosis and cancer. Best Pract Res Clin Haematol 2009; 22 (01) 61-69
    CrossrefPubMedGoogle Scholar
  • 10 Scharovsky OG, Mainetti LE, Rozados VR. Metronomic chemotherapy: changing the paradigm that more is better. Curr Oncol 2009; 16 (02) 7-15
    PubMedGoogle Scholar
  • 11 Yáñez-Mó M, Siljander PR, Andreu Z. , et al. Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles 2015; 4: 27066
    PubMedGoogle Scholar
  • 12 Aharon A, Sabbah A, Ben-Shaul S. , et al. Chemotherapy administration to breast cancer patients affects extracellular vesicles thrombogenicity and function. Oncotarget 2017; 8 (38) 63265-63280
    PubMedGoogle Scholar
  • 13 Aharon A, Tamari T, Brenner B. Monocyte-derived microparticles and exosomes induce procoagulant and apoptotic effects on endothelial cells. Thromb Haemost 2008; 100 (05) 878-885
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 14 Hernández-Vargas H, Palacios J, Moreno-Bueno G. Molecular profiling of docetaxel cytotoxicity in breast cancer cells: uncoupling of aberrant mitosis and apoptosis. Oncogene 2007; 26 (20) 2902-2913
    CrossrefPubMedGoogle Scholar
  • 15 Gardiner C, Ferreira YJ, Dragovic RA, Redman CW, Sargent IL. Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis. J Extracell Vesicles 2013; 2: 2
    PubMedGoogle Scholar
  • 16 Dai X, Yue Z, Eccleston ME, Swartling J, Slater NK, Kaminski CF. Fluorescence intensity and lifetime imaging of free and micellar-encapsulated doxorubicin in living cells. Nanomedicine (Lond) 2008; 4 (01) 49-56
    CrossrefPubMedGoogle Scholar
  • 17 Issman L, Brenner B, Talmon Y, Aharon A. Cryogenic transmission electron microscopy nanostructural study of shed microparticles. PLoS One 2013; 8 (12) e83680
    CrossrefPubMedGoogle Scholar
  • 18 Elalamy I, Mahé I, Ageno W, Meyer G. Long-term treatment of cancer-associated thrombosis: the choice of the optimal anticoagulant. J Thromb Haemost 2017; 15 (05) 848-857
    CrossrefPubMedGoogle Scholar
  • 19 Weaver BA. How Taxol/paclitaxel kills cancer cells. Mol Biol Cell 2014; 25 (18) 2677-2681
    CrossrefPubMedGoogle Scholar
  • 20 Paridaens R, Biganzoli L, Bruning P. , et al. Paclitaxel versus doxorubicin as first-line single-agent chemotherapy for metastatic breast cancer: a European Organization for Research and Treatment of Cancer Randomized Study with cross-over. J Clin Oncol 2000; 18 (04) 724-733
    CrossrefPubMedGoogle Scholar
  • 21 Owens III AP, Mackman N. Microparticles in hemostasis and thrombosis. Circ Res 2011; 108 (10) 1284-1297
    CrossrefPubMedGoogle Scholar
  • 22 Katzenell S, Shomer E, Zipori Y, Zylberfisz A, Brenner B, Aharon A. Characterization of negatively charged phospholipids and cell origin of microparticles in women with gestational vascular complications. Thromb Res 2012; 130 (03) 479-484
    CrossrefPubMedGoogle Scholar
  • 23 Shet AS, Aras O, Gupta K. , et al. Sickle blood contains tissue factor-positive microparticles derived from endothelial cells and monocytes. Blood 2003; 102 (07) 2678-2683
    CrossrefPubMedGoogle Scholar
  • 24 Connor DE, Exner T, Ma DD, Joseph JE. The majority of circulating platelet-derived microparticles fail to bind annexin V, lack phospholipid-dependent procoagulant activity and demonstrate greater expression of glycoprotein Ib. Thromb Haemost 2010; 103 (05) 1044-1052
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 25 Lima LG, Leal AC, Vargas G, Porto-Carreiro I, Monteiro RQ. Intercellular transfer of tissue factor via the uptake of tumor-derived microvesicles. Thromb Res 2013; 132 (04) 450-456
    CrossrefPubMedGoogle Scholar
  • 26 Parry GC, Erlich JH, Carmeliet P, Luther T, Mackman N. Low levels of tissue factor are compatible with development and hemostasis in mice. J Clin Invest 1998; 101 (03) 560-569
    CrossrefPubMedGoogle Scholar
  • 27 Ma L, Francia G, Viloria-Petit A. , et al. In vitro procoagulant activity induced in endothelial cells by chemotherapy and antiangiogenic drug combinations: modulation by lower-dose chemotherapy. Cancer Res 2005; 65 (12) 5365-5373
    CrossrefPubMedGoogle Scholar
  • 28 Tesselaar ME, Osanto S. Risk of venous thromboembolism in lung cancer. Curr Opin Pulm Med 2007; 13 (05) 362-367
    CrossrefPubMedGoogle Scholar
  • 29 Woei-A-Jin FJ, Tesselaar ME, Garcia Rodriguez P, Romijn FP, Bertina RM, Osanto S. Tissue factor-bearing microparticles and CA19.9: two players in pancreatic cancer-associated thrombosis?. Br J Cancer 2016; 115 (03) 332-338
    CrossrefPubMedGoogle Scholar
  • 30 Thaler J, Ay C, Mackman N. , et al. Microparticle-associated tissue factor activity, venous thromboembolism and mortality in pancreatic, gastric, colorectal and brain cancer patients. J Thromb Haemost 2012; 10 (07) 1363-1370
    CrossrefPubMedGoogle Scholar
  • 31 Geddings JE, Mackman N. Tumor-derived tissue factor-positive microparticles and venous thrombosis in cancer patients. Blood 2013; 122 (11) 1873-1880
    CrossrefPubMedGoogle Scholar
  • 32 Collier ME, Mah PM, Xiao Y, Maraveyas A, Ettelaie C. Microparticle-associated tissue factor is recycled by endothelial cells resulting in enhanced surface tissue factor activity. Thromb Haemost 2013; 110 (05) 966-976
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 33 Del Conde I, Shrimpton CN, Thiagarajan P, López JA. Tissue-factor-bearing microvesicles arise from lipid rafts and fuse with activated platelets to initiate coagulation. Blood 2005; 106 (05) 1604-1611
    CrossrefPubMedGoogle Scholar
  • 34 Davila M, Robles-Carrillo L, Unruh D. , et al. Microparticle association and heterogeneity of tumor-derived tissue factor in plasma: is it important for coagulation activation?. J Thromb Haemost 2014; 12 (02) 186-196
    CrossrefPubMedGoogle Scholar
  • 35 Garnier D, Magnus N, Lee TH. , et al. Cancer cells induced to express mesenchymal phenotype release exosome-like extracellular vesicles carrying tissue factor. J Biol Chem 2012; 287 (52) 43565-43572
    CrossrefPubMedGoogle Scholar