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Contortrostatin, a dimeric disintegrin from contortrix contortrix, inhibits breast cancer progression

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Abstract

We report the results of a multidisciplinary study on the inhibitory effect of a snake venom disintegrin, contortrostatin, a 13.5 kDa homodimeric protein isolated from Agkistrodon contortrix contortrix (southern copperhead) venom, on breast cancer progression. We demonstrate that contortrostatin binds to integrins and blocks the adhesion of human breast cancer cells (MDA-MB-435) to extracellular matrix (ECM) proteins including fibronectin and vitronectin, but it has no effect on adhesion of the cells to laminin and Matrigel. Contortrostatin also prevents invasion of MDA-MB-435 cells through an artificial Matrigel basement membrane. Daily local injection of contortrostatin (5 μg per mouse per day) into MDA-MB-435 tumor masses in an orthotopic xenograft nude mouse model inhibits growth of the tumor by 74% (p = 0.0164). More importantly, it reduces the number of pulmonary macro-metastasis of the breast cancer by 68% (p < 0.001), and micro-metastasis by 62.4% (p < 0.001). Contortrostatin is not cytotoxic to cancer cells, and does not inhibit proliferation of the breast cancer cells in vitro. However, contortrostatin inhibits angiogenesis induced by the breast cancer, as shown by immunohistochemical quantitation of the vascular endothelial cells in tumor tissue removed from the nude mice. We have identified αvβ3, an important integrin mediating cell motility and tumor invasion, as one of the binding sites of contortrostatin on MDA-MB-435 cells. We conclude that contortrostatin blocks αvβ3, and perhaps other integrins, and thus inhibits in vivo progression.

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References

  1. Hynes RO: Integrins: Versatility modulation, and signaling in cell adhesion. Cell 69: 11–25, 1992

    Google Scholar 

  2. Cheresh DA: Structural and biologic properties of integrinmediated cell adhesion. Clin Lab Med 12: 217–236, 1992

    Google Scholar 

  3. Giancotti FG, Mainiero F: Integrin-mediated adhesion and signaling in tumorgenesis. Biochim Biophys Acta 1198: 47–64, 1994

    Google Scholar 

  4. Juliano RL, Varner JA: Adhesion molecules in cancer: the role of integrins. Curr Opin Cell Biol 5: 812–818, 1993

    Google Scholar 

  5. Juliano R: Signal transduction by integrins and its role in the regulation of tumor growth. Cancer Metastasis Rev 13: 25–30, 1994

    Google Scholar 

  6. Jacob K, Bosserhoff AK, Wach F, Knuchel R, Klein EC, Hein R, Buettner R: Characterization of selected strongly and weakly invasive sublines of a primary human melanoma cell line and isolation of subtractive cDNA clones. Int J Cancer 60: 668–675, 1995

    Google Scholar 

  7. Kawahara E, Imai K, Kumagai S, Yamamoto E, Nakanishi I: Inhibitory effects of adhesion oligopeptides on the invasion of squamous carcinoma cells with special reference to implication of av integrins. J Cancer Res Clin Oncol 121: 133–140, 1995

    Google Scholar 

  8. Liapis H, Adler LM, Wick MR, Rader JS: Expression of αvβ3 integrin is less frequent in ovarian epithelial tumors of low malignant potential in contrast to ovarian carcinomas. Hum Pathol 28: 443–449, 1997

    Google Scholar 

  9. Huhtala P, Humphries MJ, McCarthy JB, Tremble PM, Werb Z, Damsky C: Cooperative signaling by α5β1 and α4β1 integrins regulates metalloproteinase gene expression in fibroblasts adhering to fibronectin. J Cell Biol 129: 867–879, 1995

    Google Scholar 

  10. Seftor REB, Seftor EA, Gehlsen KR, Stetler-Stevenson WG, Brown PD, Ruoslahti E, Hendrix MJC: Role of the αvβ3 integrin in human melanoma cell invasion. Proc Natl Acad Sci 89: 1557–1561, 1992

    Google Scholar 

  11. Seftor REB, Seftor EA, Stetler-Stevenson WG, Hendrix MJC: The 72 kDa Type IV collagenase is modulated via differential expression of αvβ3 and α5β1 integrins during human melanoma cell invasion. Cancer Res 53: 3411–3415, 1993

    Google Scholar 

  12. Brooks PC, Stromblad S, Klemke R, Visscher D, Sarkar FH, Cheresh DA: Antiintegrin αvβ3 blocks human breast cancer growth and angiogenesis in human skin. J Clin Invest 96: 1815–1822, 1995

    Google Scholar 

  13. Brooks PC, Clark RA, Cheresh DA: Requirement of vascular integrin αvβ3 for angiogenesis. Science 264: 569–571, 1994

    Google Scholar 

  14. Brooks PC, Montgomery AMP, Rosenfeld M, Reisfeld RA, Hu T, Klier G, Cheresh DA: Integrin αvβ3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels. Cell 79: 1157–1164, 1994

    Google Scholar 

  15. Pierschbacher MD, Rouslahti E: The cell attachment activity of fibronectin can be duplicated by small fragments of the molecule. Nature 309: 30–35, 1984

    Google Scholar 

  16. Knudsen KA, Tuszynski GP, Huang TF, Niewiarowski S: Trigramin, an RGD-containing peptide from snake venom, inhibits cell-substratum adhesion of human melanoma cells. Exp Cell Res 179: 42–49, 1988

    Google Scholar 

  17. Trikha M, De Clerk YA, Markland FS: Contortrostatin, a snake venom disintegrin, inhibits α1 integrin-mediated human metastatic melanoma cell adhesion, and blocks experimental metastasis. Cancer Res 54: 4993–4998, 1994

    Google Scholar 

  18. Savage B, Marzec UM, Chao BH, Harker LA, Maraganore JM, Ruggeri ZM: Binding of the snake venom-derived proteins applaggin and echistatin to the arginine- glycine- aspartic acid and recognition site(s) on platelet glycoprotein IIb/IIIa. J Biol Chem 268: 1066–1073, 1990

    Google Scholar 

  19. Juliano D, Wang Y, Marcinkiewicz C, Rosenthal LA, Stewart GJ, Niewiarowski S: Disintegrin interaction with αvβ3 integrin on human umbilical vein endothelial cells: Expression of ligand-induced binding site on α3 subunit. Exp Cell Res 225: 132–142, 1996

    Google Scholar 

  20. Dennis MS, Henzel WJ, Pitti RM, Lipari MT, Napier MA, Deisher TA, Bunting S, Lazarus RA: Platelet glycoprotein IIb/IIIa protein antagonists from snake venoms: Evidence for a family of platelet-aggregation inhibitors. Proc Natl Acad Sci 87: 2471–2475, 1990

    Google Scholar 

  21. Gould RJ, Polokoff MA, Friedman PA, Huang T-F, Holt JC, Cook JJ, Niewiarowski S: Disintegrins: A family of integrin inhibitory proteins from viper venoms. Proc Soc Exper Biol Med 195: 168–171, 1990

    Google Scholar 

  22. Niewiarowski S, McLane MA, Kloczewiak M, Stewart GJ: Disintegrins and other naturally occurring antagonists of platelet fibrinogen receptors. Semin Hematol 31: 289–300, 1994

    Google Scholar 

  23. McLane M, Marcinkiewica C, Vijay-Kumar S, Wierzbicka-Patynowski I, Niewiarowski S: Viper venom disintegrins and related molecules [Review]. Proc Soc Exp Biol Med 219: 109–119, 1998

    Google Scholar 

  24. Trikha M, Rote WE, Manley PJ, Lucchesi BR, Markland FS: Purification and characterization of platelet aggregation inhibitors from snake venoms. Thromb Res 73: 39–52, 1994

    Google Scholar 

  25. Sheu J-R, Teng C-M, Huang T-F: Triflavin, an RGDcontaining antiplatelet peptide, binds to GPIIIa of ADPstimulated platelets. Biochem Biophys Res Commun 189: 1236–1242, 1992

    Google Scholar 

  26. Kang I-C, Lee Y-D, Kim D-S: A Novel disintegrin salmosin inhibits tumor angiogenesis. Cancer Res 59: 3754–3760, 1999

    Google Scholar 

  27. Price JE, Polyzos A, Zhang RD, Daniels LM: Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice. Cancer Res 50: 717–721, 1990

    Google Scholar 

  28. Repesh LA: A new in vitro assay for quantitating tumor cell invasion. Invasion Metastasis 9: 192–208, 1989

    Google Scholar 

  29. Osborne CK, Hobbs K, Clark GM: Effect of estrogens and antiestrogens on growth of human breast cancer cells in athymic nude mice. Cancer Res 45: 584–590, 1985

    Google Scholar 

  30. Gasparini G, Harris AL: Prognostic significance of tumor vascularity. In: Teicher BA (ed.) Antiangiogenic Agents in Cancer Therapy, Totowa, Humana Press Inc., 1999, pp. 317–339

    Google Scholar 

  31. Long JS: Hypothesis testing and goodness of fit. In: Long JS (ed.) Regression Models for Categorical and Limited Dependent Variables, Thousand Oaks, SAGE, 1997, pp. 85–93

    Google Scholar 

  32. Coller BS, Peerschke EI, Scudder LE, Sullivan CA: A murine monoclonal antibody that completely blocks the binding of fibrinogen to platelets produces a thrombasthenic-like state in normal platelets and binds to glycoproteins IIb and/or IIIa. J Clin Invest 72: 325–338, 1983

    Google Scholar 

  33. Charo IF, Bekeart LS, Phillips DR: Platelet glycoprotein IIb-IIIa like proteins mediate endothelial cell attachment to adhesive proteins and the extracellular matrix. J Biol Chem 262: 9935–9938, 1987

    Google Scholar 

  34. Phillips DR, Charo IF, Scarborough RM: GP IIb-IIIa: The responsive integrin. Cell 65: 359–362, 1991

    Google Scholar 

  35. Chang YS, Chen YQ, Timar J, Nelson KK, Grossi IM, Fitzgerald LA, Diglio CA, Honn KV: Increased expression of aIIbb3 integrin in subpopulations of murine melanoma cells with high lung-colonizing ability. Int J Cancer 51: 445–451, 1992

    Google Scholar 

  36. Chen YQ, Gao X, Timar J, Tang D, Grossi IM, Chelladurai M, Kunicki TJ, Fligiel SEG, Taylor JD, Honn KV: Identification of the αIIbβ3 integrin in murine tumor cells. J Biol Chem 267: 17314–17320, 1992

    Google Scholar 

  37. Peerschke EI, Coller BS: A murine monoclonal antibody that blocks fibrinogen binding to normal platelets also inhibits fibrinogen interactions with chymotrypsin-treated platelets. Blood 64: 59–63, 1984

    Google Scholar 

  38. Liotta LA: Tumor invasion and metastasis - Role of the extracellular matrix: Rhoads Memorial Award Lecture. Cancer Res 46: 1–7, 1986

    Google Scholar 

  39. Felding-Habermann B, Cheresh DA: Vitronectin and its receptors. Curr Opin Cell Biol 5: 864–868, 1993

    Google Scholar 

  40. Wayner EA, Orlando RA, Cheresh DA: Integrins αvβ3 and αvβ5 contribute to cell attachment to vitronectin but differentially distribute on the cell surface. J Cell Biol 113: 919–929, 1991

    Google Scholar 

  41. Kleinman HK, McGarvey ML, Liotta LA, Robey PG, Tryggvason K, Martin GR: Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry 21: 6188–6193, 1982

    Google Scholar 

  42. Wong N, Mueller B, Barbas C, Ruminski P, Quaranta V, Lin E, Smith J: αv Integrins mediate adhesion and migration of breast carcinoma cell lines. Clin Exp Metastasis 16: 50–61, 1998

    Google Scholar 

  43. Klemke RL, Cai S, Giannini AL, Gallagher PJ, de Lanerolle P, Cheresh DA: Regulation of cell motility by mitogen-activated protein kinase. J Cell Biol 137: 481–492, 1997

    Google Scholar 

  44. Yeh C, Peng H, Huang T: Accutin, a new disintegrin, inhibits angiogenesis in vitro and in vivo by acting as integrin αvβ3 antagonist and inducing apoptosis. Blood 92: 3268–3276, 1998

    Google Scholar 

  45. Folkman J: Editorial: Angiogenesis and breast cancer. J Clin Oncol 12: 441–443, 1994

    Google Scholar 

  46. O'Reilly MS, Holmgren L, Chen C, Folkman J: Angiostatin induces and sustains dormancy of human primary tumors in mice. Nat Med 2: 689–692, 1996

    Google Scholar 

  47. O'Reilly MS, Boehm T, Shing Y, Fukai N, Vasios G, Lane WS, Flynn ERBJ, Olsen BR, Folkman J: Endostatin: An endogenous inhibitor of angiogenesis and tumor growth. Cell 88: 277–285, 1997

    Google Scholar 

  48. Boehm T, Folkman J, Browder T, O'Reilly MS: Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 390: 404–407, 1997

    Google Scholar 

  49. Honn KV, Tang DG, Chen YQ: Platelets and cancer metastasis: more than an epiphenomenon. Semin Thrombos Hemostas 18: 392–415, 1992

    Google Scholar 

  50. Boukerche H, Berthier-Vergnes O, Tabone E, Dore JF, Laung CL, McGregor JL: Platelet- melanoma cell interaction is mediated by the glycoprotein IIb-IIIa complex. Blood 74: 658–663, 1989

    Google Scholar 

  51. Karpatkin S, Pearlstein E, Ambrogio C, Coller BS: Role of adhesive proteins in platelet tumor interaction In Vitro and metastasis formation In Vivo. J Clin Invest 81: 1012–1019, 1988

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Los Angeles, CA, USA

    Qing Zhou, Catherine Parrish & Francis S. Markland

  2. Department of Pathology, Los Angeles, CA, USA

    Russel P. Sherwin & Valda Richters

  3. Department of Preventive Medicine, University of Southern California, Keck School of Medicine/Norris Comprehensive Cancer Center, Los Angeles, CA, USA

    Susan G. Groshen & Denice Tsao-Wei

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  1. Qing Zhou
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  2. Russel P. Sherwin
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  7. Francis S. Markland
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Zhou, Q., Sherwin, R.P., Parrish, C. et al. Contortrostatin, a dimeric disintegrin from contortrix contortrix, inhibits breast cancer progression. Breast Cancer Res Treat 61, 249–259 (2000). https://doi.org/10.1023/A:1006457903545

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