Skip to main content

Advertisement

SpringerLink
Log in

Inhibition of metastasis in a murine 4T1 breast cancer model by liposomes preventing tumor cell-platelet interactions

  • Research paper
  • Published:
Clinical & Experimental Metastasis Aims and scope Submit manuscript

Abstract

The interaction between circulating tumor cells and blood components, mainly platelets, plays an important role during metastasis. In this study, we prepared liposomes containing the platelet aggregation inhibitor Cilostazol (Cil-L). The objective of this study was to investigate the effect of this Cil-L on platelet aggregation and complex formation with murine 4T1 breast cancer cells in vitro and to determine their anti-metastatic potency in a spontaneous metastasis model of 4T1 breast cancer. Cil-L significantly inhibited the aggregation of platelets by up to 78% and completely abolished the complex formation of 4T1 tumor cells in the presence of activated platelets in vitro. Intravenous (i.v.) injection of Cil-L into mice significantly reduced the aggregability of mouse platelets by 60% measured ex vivo. To gain deeper insight into the mode of metastasis formation in a spontaneous metastasis model, 4T1 breast cancer cells were transplanted into the mammary fad pad of mice and metastasis to the mouse lungs was investigated with regard to tumor cell settlement and metastatic growth. We could demonstrate that the formation of pulmonary metastases was significantly reduced by 55% when mice were treated intravenously with 100 nmol Cil-L 6 h before tumor cell inoculation and then daily for 2 weeks. We conclude that Cil-L reduced metastasis by restricting the aggregability of mouse platelets, which probably prevents the interaction between circulating 4T1 tumor cells and platelets, making the Cil-L a useful tool for the inhibition of breast cancer metastasis in mice.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

aPlt + Cil-L:

Activated platelets + Cilostazol-liposomes

aPlt:

Activated platelets

cAMP:

Cyclic adenosine monophosphate

CH:

Cholesterol

Cil-L:

Cilostazol-encapsulating liposomes

Con-L:

Control liposomes containing OPP

DCP:

Dicetylphosphate

HPTLC:

High performance thin layer chromatographie

NaCl:

Sodiumchloride

OPP:

Perifosine

PC:

Phosphatidylcholine

PEG-PE:

N-Methyl-(polyethyleneglycyl)2000-1,2-distearyl-s,n-glycero-3-phosphatidylethanolamine

PI:

Polydispersity index

Plt:

Platelets

SD:

Standard deviation

References

  1. Tao K, Fang M, Alroy J et al (2008) Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer 8:228

    Article  PubMed  Google Scholar 

  2. Al-Mehdi AB, Tozawa K, Fisher AB et al (2000) Intravascular origin of metastasis from the proliferation of endothelium-attached tumor cells: a new model for metastasis. Nat Med 6:100–102

    Article  CAS  PubMed  Google Scholar 

  3. Monzavi-Karbassi B, Stanley JS, Hennings L et al (2007) Chondroitin sulfate glycosaminoglycans as major P-selectin ligands on metastatic breast cancer cell lines. Int J Cancer 120:1179–1191

    Article  CAS  PubMed  Google Scholar 

  4. Trousseau A (1865) Phlegmasia alba dolens. In:Clinique Medicale de l’Hotel Dieu de Paris, vol 3. JB Balliere et Fils, Paris, pp 654–712

  5. Borsig L (2008) The role of platelet activation in tumor metastasis. Expert Rev Anticancer Ther 8:1247–1255

    Article  CAS  PubMed  Google Scholar 

  6. Chen M, Geng JG (2006) P-selectin mediates adhesion of leukocytes, platelets, and cancer cells in inflammation, thrombosis, and cancer growth and metastasis. Arch Immunol Ther Exp (Warsz) 54:75–84

    Article  CAS  Google Scholar 

  7. Borsig L, Wong R, Feramisco J et al (2001) Heparin and cancer revisited: mechanistic connections involving platelets, P-selectin, carcinoma mucins, and tumor metastasis. Proc Natl Acad Sci U S A 98:3352–3357

    Article  CAS  PubMed  Google Scholar 

  8. Tsuruo T, Fujita N (2008) Platelet aggregation in the formation of tumor metastasis. Proc Jpn Acad Ser B Phys Biol Sci 84:189–198

    Article  CAS  PubMed  Google Scholar 

  9. Honn KV, Tang DG, Crissman JD (1992) Platelets and cancer metastasis: a causal relationship? Cancer Metastasis Rev 11:325–351

    Article  CAS  PubMed  Google Scholar 

  10. Tohgo A, Tanaka NG, Ogawa H (1986) Platelet-aggregating activities of metastasizing tumor cells. IV. Effects of cell surface modification on thrombin generation, platelet aggregation and subsequent lung colonization. Invasion Metastasis 6:58–68

    CAS  PubMed  Google Scholar 

  11. Nieswandt B, Hafner M, Echtenacher B et al (1999) Lysis of tumor cells by natural killer cells in mice is impeded by platelets. Cancer Res 59:1295–1300

    CAS  PubMed  Google Scholar 

  12. Gupta GP, Massague J (2004) Platelets and metastasis revisited: a novel fatty link. J Clin Invest 114:1691–1693

    CAS  PubMed  Google Scholar 

  13. Wenzel J, Zeisig R, Haider W et al. (2009) Inhibition of pulmonary metastasis in a human MT3 breast cancer xenograft model by dual liposomes preventing intravasal fibrin clot formation. Breast Cancer Res Treat Jun 23 [Epub ahead of print]

  14. Miyake K, Yamamoto S, Iijima S (1996) Blocking adhesion of cancer cells to endothelial cell types by S. agalactiae type-specific polysaccharides. Cytotechnology 22:205–209

    Article  CAS  PubMed  Google Scholar 

  15. Wahrenbrock M, Borsig L, Le D et al (2003) Selectin-mucin interactions as a probable molecular explanation for the association of Trousseau syndrome with mucinous adenocarcinomas. J Clin Invest 112:853–862

    CAS  PubMed  Google Scholar 

  16. Zeisig R, Stahn R, Wenzel K et al (2004) Effect of sialyl Lewis X-glycoliposomes on the inhibition of E-selectin-mediated tumour cell adhesion in vitro. Biochim Biophys Acta 1660:31–40

    Article  CAS  PubMed  Google Scholar 

  17. Wenzel J, Zeisig R, Fichtner I (2009) Inhibition of breast cancer metastasis by dual liposomes to disturb complex formation. Int J Pharm 370:121–128

    Article  CAS  PubMed  Google Scholar 

  18. Mancuso A, Sternberg CN (2006) New treatment approaches in metastatic renal cell carcinoma. Curr Opin Urol 16:337–341

    Article  PubMed  Google Scholar 

  19. Bobek V, Kovarik J (2004) Antitumor and antimetastatic effect of warfarin and heparins. Biomed Pharmacother 58:213–219

    Article  CAS  PubMed  Google Scholar 

  20. Hejna M, Raderer M, Zielinski CC (1999) Inhibition of metastases by anticoagulants. J Natl Cancer Inst 91:22–36

    Article  CAS  PubMed  Google Scholar 

  21. Kragh M, Loechel F (2005) Non-anti-coagulant heparins: a promising approach for prevention of tumor metastasis (review). Int J Oncol 27:1159–1167

    CAS  PubMed  Google Scholar 

  22. Borsig L, Wong R, Hynes RO et al (2002) Synergistic effects of L- and P-selectin in facilitating tumor metastasis can involve non-mucin ligands and implicate leukocytes as enhancers of metastasis. Proc Natl Acad Sci U S A 99:193–198

    Article  Google Scholar 

  23. Yoshimoto T, Shirasaka T, Fujimoto S et al (2009) Cilostazol may prevent cerebral vasospasm following subarachnoid hemorrhage. Neurol Med Chir (Tokyo) 49:235–240

    Article  Google Scholar 

  24. Woo SK, Kang WK, Kwon KI (2002) Pharmacokinetic and pharmacodynamic modeling of the antiplatelet and cardiovascular effects of cilostazol in healthy humans. Clin Pharmacol Ther 71:246–252

    Article  CAS  PubMed  Google Scholar 

  25. Uehara S, Hirayama A (1989) Effects of cilostazol on platelet function. Arzneimittelforschung 39:1531–1534

    CAS  PubMed  Google Scholar 

  26. Pulaski BA, Clements VK, Pipeling MR et al (2000) Immunotherapy with vaccines combining MHC class II/CD80+ tumor cells with interleukin-12 reduces established metastatic disease and stimulates immune effectors and monokine induced by interferon gamma. Cancer Immunol Immunother 49:34–45

    Article  CAS  PubMed  Google Scholar 

  27. Morecki S, Yacovlev L, Slavin S (1998) Effect of indomethacin on tumorigenicity and immunity induction in a murine model of mammary carcinoma. Int J Cancer 75:894–899

    Article  CAS  PubMed  Google Scholar 

  28. Heppner GH, Miller FR, Shekhar PM (2000) Nontransgenic models of breast cancer. Breast Cancer Res 2:331–334

    Article  CAS  PubMed  Google Scholar 

  29. Zeisig R, Muller K, Maurer N et al (2001) The composition-dependent presence of free (micellar) alkylphospholipid in liposomal formulations of octadecyl-1, 1-dimethyl-piperidino-4-yl-phosphate affects its cytotoxic activity in vitro. J Membr Biol 182:61–69

    CAS  PubMed  Google Scholar 

  30. Naundorf H, Rewasowa EC, Fichtner I et al (1992) Characterization of two human mammary carcinomas, MT-1 and MT-3, suitable for in vivo testing of ether lipids and their derivatives. Breast Cancer Res Treat 23:87–95

    Article  CAS  PubMed  Google Scholar 

  31. Zeisig R, Arndt D, Stahn R et al (1998) Physical properties and pharmacological activity in vitro and in vivo of optimised liposomes prepared from a new cancerostatic alkylphospholipid. Biochim Biophys Acta 1414:238–248

    Article  CAS  PubMed  Google Scholar 

  32. Cardinal DC, Flower RJ (1980) The electronic aggregometer: a novel device for assessing platelet behavior in blood. J Pharmacol Methods 3:135–158

    Article  CAS  PubMed  Google Scholar 

  33. Minami N, Suzuki Y, Yamamoto M et al (1997) Inhibition of shear stress-induced platelet aggregation by cilostazol, a specific inhibitor of cGMP-inhibited phosphodiesterase, in vitro and ex vivo. Life Sci 61(25):383–389

    Article  Google Scholar 

  34. Im JH, Fu W, Wang H et al (2004) Coagulation facilitates tumor cell spreading in the pulmonary vasculature during early metastatic colony formation. Cancer Res 64:8613–8619

    Article  CAS  PubMed  Google Scholar 

  35. Saiki I, Koike C, Obata A et al (1996) Functional role of sialyl Lewis X and fibronectin-derived RGDS peptide analogue on tumor-cell arrest in lungs followed by extravasation. Int J Cancer 65:833–839

    Article  CAS  PubMed  Google Scholar 

  36. Beviglia L, Stewart GJ, Niewiarowski S (1995) Effect of four disintegrins on the adhesive and metastatic properties of B16F10 melanoma cells in a murine model. Oncol Res 7:7–20

    CAS  PubMed  Google Scholar 

  37. Oleksowicz L, Mrowiec Z, Schwartz E et al (1995) Characterization of tumor-induced platelet aggregation: the role of immunorelated GPIb and GPIIb/IIIa expression by MCF-7 breast cancer cells. Thromb Res 79:261–274

    Article  CAS  PubMed  Google Scholar 

  38. Gasic GJ (1984) Role of plasma, platelets, and endothelial cells in tumor metastasis. Cancer Metastasis Rev 3:99–114

    Article  CAS  PubMed  Google Scholar 

  39. Price JT, Thompson EW (2002) Mechanisms of tumour invasion and metastasis: emerging targets for therapy. Expert Opin Ther Targets 6:217–233

    Article  CAS  PubMed  Google Scholar 

  40. Dawson DL, Cutler BS, Meissner MH et al (1998) Cilostazol has beneficial effects in treatment of intermittent claudication: results from a multicenter, randomized, prospective, double-blind trial. Circulation 98:678–686

    CAS  PubMed  Google Scholar 

  41. Wong CW, Song C, Grimes MM et al (2002) Intravascular location of breast cancer cells after spontaneous metastasis to the lung. Am J Pathol 161:749–753

    PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the Federal Ministry of Education and Research of Germany (Biochance PLUS program; PTJ-BIO/0313601). We thank Prof. M. van der Giet for supporting the PhD work of J.W. at the Charite Berlin; M. Becker (MDC Berlin-Buch) for the excellent performance of the animal experiments; and Lipoid GmbH Ludwigshafen for providing us with egg phosphatidylcholine.

Conflict of interest statement

All authors confirm that they have no potential conflict of interest, including any financial, personal, or other relationships with other people or organizations within that could inappropriately influence (bias) their work.

Author information

Authors and Affiliations

  1. Max-Delbrück-Center for Molecular Medicine Berlin-Buch, R.-Rössle-Strasse 10, 13125, Berlin, Germany

    Jane Wenzel & Iduna Fichtner

  2. Experimental Pharmacology and Oncology, Berlin-Buch GmbH (EPO GmbH), R.-Rössle-Strasse 10, 13125, Berlin, Germany

    Reiner Zeisig

  3. Charité, Medical Clinic IV, Nephrology, Campus Benjamin Franklin, Berlin, Germany

    Jane Wenzel

Authors
  1. Jane Wenzel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Reiner Zeisig
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Iduna Fichtner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jane Wenzel.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wenzel, J., Zeisig, R. & Fichtner, I. Inhibition of metastasis in a murine 4T1 breast cancer model by liposomes preventing tumor cell-platelet interactions. Clin Exp Metastasis 27, 25–34 (2010). https://doi.org/10.1007/s10585-009-9299-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10585-009-9299-y

Keywords

Search

Navigation