Antimigratory and antimetastatic effect of heparin-derived 4–18 unit oligosaccharides in a preclinical human melanoma metastasis model

Summary

Heparin and its derivatives have been shown to inhibit angiogenesis and metastasis formation. Accordingly, we investigated the effect of heparin fragments containing 4 to 22 monomers on human melanoma cell proliferation, migration and invasion in vitro as well as on the in vivo metastatic potential in a SCID mouse model. Only oligosaccharide dp18 had significant inhibitory effect on cell proliferation. In contrast, cell migration was inhibited by all oligosaccharides studied except dp8 and dp22. Anti CD44v3 antibody stimulated cell migration and invasion, and this effect could be attenuated by oligosaccharides dp4 and dp18. These fragments also inhibited the catalytic activity of myosin light chain phosphatase as well. Moreover, oligosaccharides dp4 and dp18 reduced the number of lung colonies formed in SCID mice intravenously injected with human melanoma cells, while dp22 proved to be ineffective in this respect.These studies revealed that fragments of heparin have an antimigratory and antimetastatic potential. These fragments lack the haemostatic effect of heparin, suggesting that they are potential specific antimetastatic agents in anticancer therapy.

Keywords

Melanoma - oligosaccharides - cell - migration - xenograft - metastasis model

• References

• 1 Lip GY, Chin BS, Blann AD. Cancer and the prothrombotic state. Lancet Oncol 2002; 03: 27-34.
• 2 Gieseler F, Luhr I, Kunze T. et al. Activated coagulation factors in human malignant effusions and their contribution to cancer cell metastasis and therapy. Thromb Haemost 2007; 97: 1023-1030.
• 3 Seddighzadeh A, Shetty R, Goldhaber SZ. Venous thromboembolism in patients with active cancer. Thromb Haemost 2007; 98: 656-661.
• 4 Zacharski LR, Ornstein DL. Heparin and cancer. Thromb Haemost 1998; 80: 10-23.
• 5 Lebeau B, Chastang C, Brechot JM. et al. Subcutaneous heparin treatment increases survival in small cell lung cancer. “Petites Cellules” Group. Cancer 1994; 74: 38-45.
• 6 von Tempelhoff GF, Harenberg J, Niemann F. et al. Effect of low molecular weight heparin (Certoparin) versus unfractionated heparin on cancer survival following breast and pelvic cancer surgery: A prospective randomized double-blind trial. Int J Oncol 2000; 16: 815-824.
• 7 Constantini S, Kanner A, Friedman A. et al. Safety of perioperative minidose heparin in patients undergoing brain tumor surgery: a prospective, randomized, double-blind study. J Neurosurg 2001; 94: 918-921.
• 8 Lindahl U. Heparan sulfate-protein interactions--a concept for drug design?. Thromb Haemost 2007; 98: 109-115.
• 9 Timar J, Tovari J, Raso E. et al. Platelet-mimicry of cancer cells: epiphenomenon with clinical significance. Oncology 2005; 69: 185-201.
• 10 Smorenburg SM, Van Noorden CJ. The complex effects of heparins on cancer progression and metastasis in experimental studies. Pharmacol Rev 2001; 53: 93-105.
• 11 Sciumbata T, Caretto P, Pirovano P. et al. Treatment with modified heparins inhibits experimental metastasis formation and leads, in some animals, to long-term survival. Invasion Metastasis 1996; 16: 132-143.
• 12 Amirkhosravi A, Mousa SA, Amaya M. et al. Antimetastatic effect of tinzaparin, a low-molecular-weight heparin. J Thromb Haemost 2003; 01: 1972-1976.
• 13 Bereczky B, Gilly R, Raso E. et al. Selective antimetastatic effect of heparins in preclinical human melanoma models is based on inhibition of migration and microvascular arrest. Clin Exp Metastasis 2005; 22: 69-76.
• 14 Sutton A, Friand V, Papy-Garcia D. et al. Glycosaminoglycans and their synthetic mimetics inhibit RANTES-induced migration and invasion of human hepatoma cells. Mol Cancer Ther 2007; 06: 2948-2958.
• 15 Hasan J, Shnyder SD, Clamp AR. et al. Heparin octasaccharides inhibit angiogenesis in vivo. Clin Cancer Res 2005; 11: 8172-8179.
• 16 Griffioen AW, Coenen MJ, Damen CA. et al. CD44 is involved in tumor angiogenesis; an activation antigen on human endothelial cells. Blood 1997; 90: 1150-1159.
• 17 Bennett KL, Jackson DG, Simon JC. et al. CD44 isoforms containing exon V3 are responsible for the presentation of heparin-binding growth factor. J Cell Biol 1995; 128: 687-698.
• 18 Forster-Horvath C, Meszaros L, Raso E. et al. Expression of CD44v3 protein in human endothelial cells in vitro and in tumoral microvessels in vivo. Microvasc Res 2004; 68: 110-118.
• 19 Dome B, Somlai B, Ladanyi A. et al. Expression of CD44v3 splice variant is associated with the visceral metastatic phenotype of human melanoma. Virchows Arch 2001; 439: 628-635.
• 20 Ladanyi A, Timar J, Paku S. et al. Selection and characterization of human melanoma lines with different liver-colonizing capacity. Int J Cancer 1990; 46: 456-461.
• 21 Rai-el-Balhaa G, Pellerin JL, Bodin G. et al. Lymphoblastic transformation assay of sheep peripheral blood lymphocytes: a new rapid and easy-to-read technique. Comp Immunol Microbiol Infect Dis 1985; 08: 311-318.
• 22 Albini A, Iwamoto Y, Kleinman HK. et al. A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 1987; 47: 3239-3245.
• 23 Gergely P, Erdodi F, Bot G. Heparin inhibits the activity of protein phosphatase-1. FEBS Lett 1984; 169: 45-48.
• 24 Herault JP, Bernat A, Roye F. et al. Pharmacokinetics of new synthetic heparin mimetics. Thromb Haemost 2002; 87: 985-989.
• 25 Sasisekharan R, Venkataraman G. Heparin and heparan sulfate: biosynthesis, structure and function. Curr Opin Chem Biol 2000; 04: 626-631.
• 26 Gallagher JT. Heparan sulfate: growth control with a restricted sequence menu. J Clin Invest 2001; 108: 357-361.
• 27 Mellor P, Harvey JR, Murphy KJ. et al. Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDAMB 231 breast cancer cells in vivo. Br J Cancer 2007; 97: 761-768.
• 28 Burger JA, Kipps TJ. CXCR4: a key receptor in the crosstalk between tumor cells and their microenvironment. Blood 2006; 107: 1761-1767.
• 29 Seiter S, Schadendorf D, Herrmann K. et al. Expression of CD44 variant isoforms in malignant melanoma. Clin Cancer Res 1996; 02: 447-456.
• 30 Manten-Horst E, Danen EH, Smit L. et al. Expression of CD44 splice variants in human cutaneous melanoma and melanoma cell lines is related to tumor progression and metastatic potential. Int J Cancer 1995; 64: 182-188.
• 31 Lesley J, Hyman R. CD44 structure and function. Front Biosci 1998; 03: d616-30.
• 32 Fazekas K, Raso E, Zarandi M. et al. Basic HGFlike peptides inhibit generation of liver metastases in murine and human tumor models. Anticancer Res 2002; 22: 2575-2579.
• 33 van der Voort R, Taher TE, Wielenga VJ. et al. Heparan sulfate-modified CD44 promotes hepatocyte growth factor/scatter factor-induced signal transduction through the receptor tyrosine kinase c-Met. J Biol Chem 1999; 274: 6499-6506.
• 34 Jackson DG, Bell JI, Dickinson R. et al. Proteoglycan forms of the lymphocyte homing receptor CD44 are alternatively spliced variants containing the v3 exon. J Cell Biol 1995; 128: 673-685.
• 35 Natali PG, Nicotra MR, Di Renzo MF. et al. Expression of the c-Met/HGF receptor in human melanocytic neoplasms: demonstration of the relationship to malignant melanoma tumour progression. Br J Cancer 1993; 68: 746-750.
• 36 Borsig L, Wong R, Feramisco J. et al. Heparin and cancer revisited: mechanistic connections involving platelets, P-selectin, carcinoma mucins, and tumor metastasis. Proc Natl Acad Sci USA 2001; 98: 3352-3357.
• 37 Alves CS, Burdick MM, Thomas SN. et al. The dual role of CD44 as a functional P-selectin ligand and fibrin receptor in colon carcinoma cell adhesion. Am J Physiol Cell Physiol 2008; 294: C907-916.
• 38 Napier SL, Healy ZR, Schnaar RL. et al. Selectin ligand expression regulates the initial vascular interactions of colon carcinoma cells: the roles of CD44v and alternative sialofucosylated selectin ligands. J Biol Chem 2007; 282: 3433-3441.
• 39 Borsig L, Wong R, Hynes RO. et al. 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 USA 2002; 99: 2193-2198.
• 40 Thomas SN, Schnaar RL, Konstantopoulos K. Podocalyxin-like protein is an E-/L-selectin ligand on colon carcinoma cells: comparative biochemical properties of selectin ligands in host and tumor cells. Am J Physiol Cell Physiol 2009; 296: C505-513.
• 41 Ludwig RJ, Boehme B, Podda M. et al. Endothelial P-selectin as a target of heparin action in experimental melanoma lung metastasis. Cancer Res 2004; 64: 2743-2750.
• 42 Fritzsche J, Simonis D, Bendas G. Melanoma cell adhesion can be blocked by heparin in vitro: suggestion of VLA-4 as a novel target for antimetastatic approaches. Thromb Haemost 2008; 100: 1166-1175.
• 43 Stevenson JL, Varki A, Borsig L. Heparin attenuates metastasis mainly due to inhibition of P-and L-selectin, but non-anticoagulant heparins can have additional effects. Thromb Res 2007; 120 (Suppl. 02) S107-111.
• 44 Friedl P, Wolf K. Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 2003; 03: 362-374.
• 45 Wolf K, Mazo I, Leung H. et al. Compensation mechanism in tumor cell migration: mesenchymalamoeboid transition after blocking of pericellular proteolysis. J Cell Biol 2003; 160: 267-277.
• 46 Mahalingam Y, Gallagher JT, Couchman JR. Cellular adhesion responses to the heparin-binding (HepII) domain of fibronectin require heparan sulfate with specific properties. J Biol Chem 2007; 282: 3221-3230.
• 47 Timar J, Moczar M, Lapis K. et al. Interactions of exogenous heparan sulfate with tumor cells of different metastatic phenotype. Invasion Metastasis 1990; 10: 301-315.
• 48 Kovalszky I, Dudas J, Olah-Nagy J. et al. Inhibition of DNA topoisomerase I activity by heparan sulfate and modulation by basic fibroblast growth factor. Mol Cell Biochem 1998; 183: 11-23.
• 49 Wakatsuki T, Wysolmerski RB, Elson EL. Mechanics of cell spreading: role of myosin II. J Cell Sci 2003; 116: 1617-1625.
• 50 Hartshorne DJ, Ito M, Erdodi F. Role of protein phosphatase type 1 in contractile functions: myosin phosphatase. J Biol Chem 2004; 279: 37211-37214.
• 51 Hirano K, Derkach DN, Hirano M. et al. Protein kinase network in the regulation of phosphorylation and dephosphorylation of smooth muscle myosin light chain. Mol Cell Biochem 2003; 248: 105-114.
• 52 Somlyo AP, Somlyo AV. Signal transduction by G-proteins, rho-kinase and protein phosphatase to smooth muscle and non-muscle myosin II. J Physiol 2000; 522: 177-185.
• 53 Yong J, Tan I, Lim L. et al. Phosphorylation of myosin phosphatase targeting subunit 3 (MYPT3) and regulation of protein phosphatase 1 by protein kinase A. J Biol Chem 2006; 281: 31202-31211.
• 54 Lontay B, Kiss A, Gergely P. et al. Okadaic acid induces phosphorylation and translocation of myosin phosphatase target subunit 1 influencing myosin phosphorylation, stress fiber assembly and cell migration in HepG2 cells. Cell Signal 2005; 17: 1265-1275.