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Clinical & Experimental Metastasis

, Volume 29, Issue 5, pp 431–439 | Cite as

Inhibitory effect of non-anticoagulant heparin (S-NACH) on pancreatic cancer cell adhesion and metastasis in human umbilical cord vessel segment and in mouse model

  • Thangirala Sudha
  • Patricia Phillips
  • Camille Kanaan
  • Robert J. Linhardt
  • Lubor Borsig
  • Shaker A. MousaEmail author
Research Paper
First Online:

Abstract

Metastasis is the most devastating aspect of cancer and it is the main cause of morbidity and mortality in cancer patients. Tumor cell adhesion to the vascular endothelial cell lining is an important step in metastatic progression and is prompted by platelets. Mucin 1 is over-expressed and aberrantly glycosylated in more than 60% of pancreatic ductal adeno-carcinomas, which mediate adhesion of pancreatic cancer cells to platelets via P-selectin. The anticoagulant low molecular weight heparins (LMWHs), which are commonly used in venous Thromboprophylaxis and treatment, appear to have an effect on cancer survival. The aim of this study is to investigate the effect of platelets on human pancreatic cancer MPanc96 cell adhesion to the endothelial cell vessel wall, and to examine the effect of heparin derivatives on MPanc96 adhesion using a novel, in vitro model of human umbilical cord vein. The modified heparin S-NACH (sulfated non-anticoagulant heparin), which is devoid of antithrombin (AT) binding and devoid of inhibition of systemic AT-dependent coagulation factors such as factor Xa and IIa, and the LMWH tinzaparin both potently reduced adhesion and invasion of fluorescence-labeled MPanc96 cancer cells to the endothelial layer of umbilical cord vein in a dose-dependent manner. S-NACH effectively inhibited P-selectin mediated MPanc96 cell adhesion, and inhibited cell adhesion and invasion similar to tinzaparin, indicating that systemic anticoagulation is not a necessary component for heparin attenuation of cancer cell adhesion, invasion, and metastasis. Also, S-NACH and tinzaparin versus unfractionated heparin, heparin derivatives enoxaparin, deltaparin, fraxiparin, and fondaparinux were evaluated for their effect on platelet-cancer cell adhesion. An in vivo anti-metastatic S-NACH-treated nude mouse model of MPanc96 pancreatic cancer cell metastasis demonstrated potent anti-metastasis efficacy as evidenced by IVIS imaging and histological staining.

Keywords

Heparin Low molecular weight heparin Anticoagulant Heparin derivatives P-selectin glycoprotein ligand-1 P-selectin Platelet Endothelial cells Cancer cell adhesion Invasion Metastasis 
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Notes

Acknowledgments

This work was supported by NIH grant R21 CA124931. We thank Dr. Howard Smith of Albany Medical College for his assistance with the IRB approval process and continuous support. We appreciate the excellent editing by Dr. Kelly Keating and the technical support by members of the Pharmaceutical Research Institute, Rensselaer, NY.

References

  1. 1.
    Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ (2007) Cancer statistics, 2007. CA Cancer J Clin 57(1):43–66PubMedCrossRefGoogle Scholar
  2. 2.
    Hidalgo M (2010) Pancreatic cancer. N Engl J Med 362(17):1605–1617. doi: 10.1056/NEJMra0901557 PubMedCrossRefGoogle Scholar
  3. 3.
    Gay LJ, Felding-Habermann B (2011) Contribution of platelets to tumour metastasis. Nat Rev Cancer 11(2):123–134. doi: 10.1038/nrc3004 PubMedCrossRefGoogle Scholar
  4. 4.
    Zhang N, Zhang WJ, Cai HQ, Liu HL, Peng L, Li CH, Ye LY, Xu SQ, Yang ZH, Lou JN (2011) Platelet adhesion and fusion to endothelial cell facilitate the metastasis of tumor cell in hypoxia-reoxygenation condition. Clin Exp Metastasis 28(1):1–12. doi: 10.1007/s10585-010-9353-9 PubMedCrossRefGoogle Scholar
  5. 5.
    Mousa SA (2004) Low-molecular-weight heparin in thrombosis and cancer. Semin Thromb Hemost 30(Suppl 1):25–30. doi: 10.1055/s-2004-823000 PubMedGoogle Scholar
  6. 6.
    Khorana AA, Fine RL (2004) Pancreatic cancer and thromboembolic disease. Lancet Oncol 5(11):655–663. doi: 10.1016/S1470-2045(04)01606-7 PubMedCrossRefGoogle Scholar
  7. 7.
    Heinmoller E, Schropp T, Kisker O, Simon B, Seitz R, Weinel RJ (1995) Tumor cell-induced platelet aggregation in vitro by human pancreatic cancer cell lines. Scand J Gastroenterol 30(10):1008–1016PubMedCrossRefGoogle Scholar
  8. 8.
    Mousa SA, Petersen LJ (2009) Anti-cancer properties of low-molecular-weight heparin: preclinical evidence. Thromb Haemost 102(2):258–267. doi: 10.1160/TH08-12-0832 PubMedGoogle Scholar
  9. 9.
    Icli F, Akbulut H, Utkan G, Yalcin B, Dincol D, Isikdogan A, Demirkazik A, Onur H, Cay F, Buyukcelik A (2007) Low molecular weight heparin (LMWH) increases the efficacy of cisplatinum plus gemcitabine combination in advanced pancreatic cancer. J Surg Oncol 95(6):507–512. doi: 10.1002/jso.20728 PubMedCrossRefGoogle Scholar
  10. 10.
    Mousa SA, Mohamed S (2004) Inhibition of endothelial cell tube formation by the low molecular weight heparin, tinzaparin, is mediated by tissue factor pathway inhibitor. Thromb Haemost 92(3):627–633. doi: 10.1267/THRO04090000 PubMedGoogle Scholar
  11. 11.
    Wahrenbrock M, Borsig L, Le D, Varki N, Varki A (2003) Selectin–mucin interactions as a probable molecular explanation for the association of Trousseau syndrome with mucinous adenocarcinomas. J Clin Invest 112(6):853–862. doi: 10.1172/JCI18882 PubMedGoogle Scholar
  12. 12.
    Hejna M, Raderer M, Zielinski CC (1999) Inhibition of metastases by anticoagulants. J Natl Cancer Inst 91(1):22–36PubMedCrossRefGoogle Scholar
  13. 13.
    Borsig L, Wong R, Feramisco J, Nadeau DR, Varki NM, Varki A (2001) Heparin and cancer revisited: mechanistic connections involving platelets, P-selectin, carcinoma mucins, and tumor metastasis. Proc Natl Acad Sci U S A 98(6):3352–3357. doi: 10.1073/pnas.061615598 PubMedCrossRefGoogle Scholar
  14. 14.
    Stevenson JL, Varki A, Borsig L (2007) Heparin attenuates metastasis mainly due to inhibition of P- and L-selectin, but non-anticoagulant heparins can have additional effects. Thromb Res 120(Suppl 2):S107–S111. doi: 10.1016/S0049-3848(07)70138-X PubMedCrossRefGoogle Scholar
  15. 15.
    Parish CR, Freeman C, Brown KJ, Francis DJ, Cowden WB (1999) Identification of sulfated oligosaccharide-based inhibitors of tumor growth and metastasis using novel in vitro assays for angiogenesis and heparanase activity. Cancer Res 59(14):3433–3441PubMedGoogle Scholar
  16. 16.
    Vlodavsky I, Mohsen M, Lider O, Svahn CM, Ekre HP, Vigoda M, Ishai-Michaeli R, Peretz T (1994) Inhibition of tumor metastasis by heparanase inhibiting species of heparin. Invasion Metastasis 14(1–6):290–302PubMedGoogle Scholar
  17. 17.
    Klerk CP, Smorenburg SM, Otten HM, Lensing AW, Prins MH, Piovella F, Prandoni P, Bos MM, Richel DJ, van Tienhoven G, Buller HR (2005) The effect of low molecular weight heparin on survival in patients with advanced malignancy. J Clin Oncol 23(10):2130–2135. doi: 10.1200/JCO.2005.03.134 PubMedCrossRefGoogle Scholar
  18. 18.
    Agnelli G, Gussoni G, Bianchini C, Verso M, Mandala M, Cavanna L, Barni S, Labianca R, Buzzi F, Scambia G, Passalacqua R, Ricci S, Gasparini G, Lorusso V, Bonizzoni E, Tonato M (2009) Nadroparin for the prevention of thromboembolic events in ambulatory patients with metastatic or locally advanced solid cancer receiving chemotherapy: a randomised, placebo-controlled, double-blind study. Lancet Oncol 10(10):943–949. doi: 10.1016/S1470-2045(09)70232-3 PubMedCrossRefGoogle Scholar
  19. 19.
    von Delius S, Ayvaz M, Wagenpfeil S, Eckel F, Schmid RM, Lersch C (2007) Effect of low-molecular-weight heparin on survival in patients with advanced pancreatic adenocarcinoma. Thromb Haemost 98(2):434–439Google Scholar
  20. 20.
    Maraveyas A, Ettelaie C, Echrish H, Li C, Gardiner E, Greenman J, Madden LA (2010) Weight-adjusted dalteparin for prevention of vascular thromboembolism in advanced pancreatic cancer patients decreases serum tissue factor and serum-mediated induction of cancer cell invasion. Blood Coagul Fibrinolysis 21(5):452–458. doi: 10.1097/MBC.0b013e328338dc49 PubMedCrossRefGoogle Scholar
  21. 21.
    Green D, Hull RD, Brant R, Pineo GF (1992) Lower mortality in cancer patients treated with low-molecular-weight versus standard heparin. Lancet 339(8807):1476PubMedCrossRefGoogle Scholar
  22. 22.
    Prandoni P, Lensing AW, Buller HR, Carta M, Cogo A, Vigo M, Casara D, Ruol A, ten Cate JW (1992) Comparison of subcutaneous low-molecular-weight heparin with intravenous standard heparin in proximal deep-vein thrombosis. Lancet 339(8791):441–445PubMedCrossRefGoogle Scholar
  23. 23.
    Gould MK, Dembitzer AD, Doyle RL, Hastie TJ, Garber AM (1999) Low-molecular-weight heparins compared with unfractionated heparin for treatment of acute deep venous thrombosis. A meta-analysis of randomized, controlled trials. Ann Intern Med 130(10):800–809PubMedGoogle Scholar
  24. 24.
    Mousa SA, Linhardt R, Francis JL, Amirkhosravi A (2006) Anti-metastatic effect of a non-anticoagulant low-molecular-weight heparin versus the standard low-molecular-weight heparin, enoxaparin. Thromb Haemost 96(6):816–821PubMedGoogle Scholar
  25. 25.
    Yoshitomi Y, Nakanishi H, Kusano Y, Munesue S, Oguri K, Tatematsu M, Yamashina I, Okayama M (2004) Inhibition of experimental lung metastases of Lewis lung carcinoma cells by chemically modified heparin with reduced anticoagulant activity. Cancer Lett 207(2):165–174. doi: 10.1016/j.canlet.2003.11.037 PubMedCrossRefGoogle Scholar
  26. 26.
    Borsig L, Wong R, Hynes RO, Varki NM, Varki A (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(4):2193–2198. doi: 10.1073/pnas.261704098 PubMedCrossRefGoogle Scholar
  27. 27.
    Sehgal LR, Wong J, He J, Wood T, Takagi I, Eldibany M, Caprini J, Mousa SA (2005) Novel in vitro perfusion model to study the interaction between coagulation and blood-borne metastasis. J Cell Biochem 96(4):700–708. doi: 10.1002/jcb.20571 PubMedCrossRefGoogle Scholar
  28. 28.
    Lapierre F, Holme K, Lam L, Tressler RJ, Storm N, Wee J, Stack RJ, Castellot J, Tyrrell DJ (1996) Chemical modifications of heparin that diminish its anticoagulant but preserve its heparanase-inhibitory, angiostatic, anti-tumor and anti-metastatic properties. Glycobiology 6(3):355–366PubMedCrossRefGoogle Scholar
  29. 29.
    Sciumbata T, Caretto P, Pirovano P, Pozzi P, Cremonesi P, Galimberti G, Leoni F, Marcucci F (1996) Treatment with modified heparins inhibits experimental metastasis formation and leads, in some animals, to long-term survival. Invasion Metastasis 16(3):132–143PubMedGoogle Scholar
  30. 30.
    Gomes N, Vassy J, Lebos C, Arbeille B, Legrand C, Fauvel-Lafeve F (2004) Breast adenocarcinoma cell adhesion to the vascular subendothelium in whole blood and under flow conditions: effects of alphavbeta3 and alphaIIbbeta3 antagonists. Clin Exp Metastasis 21(6):553–561PubMedCrossRefGoogle Scholar
  31. 31.
    Besmer DM, Curry JM, Roy LD, Tinder TL, Sahraei M, Schettini J, Hwang SI, Lee YY, Gendler SJ, Mukherjee P (2011) Pancreatic ductal adenocarcinoma mice lacking mucin 1 have a profound defect in tumor growth and metastasis. Cancer Res 71(13):4432–4442. doi: 10.1158/0008-5472.CAN-10-4439 PubMedCrossRefGoogle Scholar
  32. 32.
    Kragh M, Binderup L, Vig Hjarnaa PJ, Bramm E, Johansen KB, Frimundt Petersen C (2005) Non-anti-coagulant heparin inhibits metastasis but not primary tumor growth. Oncol Rep 14(1):99–104PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Thangirala Sudha
    • 1
  • Patricia Phillips
    • 1
  • Camille Kanaan
    • 2
  • Robert J. Linhardt
    • 3
  • Lubor Borsig
    • 4
  • Shaker A. Mousa
    • 1
    Email author
  1. 1.Pharmaceutical Research Institute at Albany College of Pharmacy and Health SciencesRensselaerUSA
  2. 2.Albany Medical Center, Obstetrics and GynecologyAlbanyUSA
  3. 3.Rensselaer Polytechnic InstituteBiotechnology CenterTroyUSA
  4. 4.Institute of PhysiologyUniversity of Zürich-IrchelZurichSwitzerland

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