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In vitro assay of the invasive potential of malignant bone and soft tissue tumours through basement membranes

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Summary

We have used reconstituted basement membrane molecules which have formed into barriers in order to investigate the invasive potential of malignant bone and soft tissue tumour cells in vitro. A number of cell lines established from human malignant tumours demonstrated a high degree of invasiveness, although fibroblasts showed no ability to penetrate the basement membrane barrier. H-ras oncogene transfected cells into the fibroblasts were much more invasive than the parent lines. Primary cultures of malignant tumour cells demonstrated invasiveness, while those of nonmetastatic cells and fibroblasts did not. The binding of tumour cells to laminin in the basement membranes was found to induce secretion of collagenase and motility which are crucial factors for invasion. A synthetic peptide, Tyr-Ile-Gly-Ser-Arg, was able to suppress the invasiveness of HT1080 human fibrosarcoma cells, and also reduced lung colonisation in vitro. The results suggest that the in vitro assay was useful, firstly to determine the invasive potential, secondly to investigate the mechanism of invasion, and finally to development treatment against invasion and metastases.

Résumé

Afin d'étudier la prolifération in vitro des cellules tumorales malignes des os et des tissus mous, nous avons utilisé des molécules reconstituées de la membrane basale organisées en barrières. Un grand nombre de cellules provenant de tumeurs malignes humaines ont montré un taux élevé de prolifération bien que les fibroblastes n'aient fait preuve d'aucune aptitude à franchir la barrière de la membrane basale. Dans les fibroblastes les cellules infectées par le H-ras oncogène ont proliféré beaucoup plus que les rangées apparentées. Les cultures primaires de cellules tumorales malignes ont montré cette prolifération, contrairement aux cellules non-métastatiques et aux fibroblastes. On a constaté que la liaison dans la membrane basale des cellules tumorales à la laminine déclenche la sécrétion de collagènase IV et la motilité, qui sont considérées comme les deux facteurs cruciaux de la prolifération. Nous avons réalisé ces études de la prolifération dans le but de tester les facteurs susceptibles de l'entraver. Un peptide synthétique, le Tyr-Ile-Gly-Ser-Arg (YIGSR), de la chaîne B1 de la laminine s'est montré capable d'arrêter la prolifération des cellules du sarcome humain HT1080, l'YIGSR peut également réduire in vivo l'envahissement du poumon. Ces résultats permettent de penser que ces expériences sont utiles, essentiellement pour déterminer le potentiel de prolifération, ensuite pour préciser le mécanisme de l'envahissement et enfin pour mettre au point des thérapeutiques efficaces contre la prolifération et les métastases des tumeurs malignes des os et des tissus mous.

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References

  1. Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM, McEwan RN (1987) A rapid in vitro invasion assay for quantitating the invasive potential of tumor cells. Cancer Res 47: 3239–3245

    CAS  PubMed  Google Scholar 

  2. Billiau A, Edy VG, Heremans H, Damme J, Van Damme J, Desmyer J, Georgiades JA, De Somer P 1977) Human interferon: Mass production in a newly established cell line, MG63. Antimicrobiol Ag Chemother 12: 11–15

    Google Scholar 

  3. Bradley MO, Kraynak AR, Storer RD, Gibbs JB (1986) Experimental metastasis in nude mice of NIH3T3 cells containing various ras genes. Proc Natl Acad Aci USA 83: 5277–5281

    Google Scholar 

  4. Fogh J, Trempe G (1975) In: Fogh J (ed) Human tumor cells in vitro. Plenum Press, New York, pp 115–159

    Google Scholar 

  5. Giard DJ, Aaronson SA, Todaro GJ, Arnstein P, Kersey JH, Dosik H, Parks WP (1973) In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51: 1417–1423

    CAS  PubMed  Google Scholar 

  6. Graf J, Iwamoto Y, Sasaki M, Martin GR, Kleinman HK, Robey FA, Yamada Y (1987) Identification of an amino acid sequence in laminin mediating cell attachment, chemotaxis and receptor binding. Cell 48: 989–996

    Google Scholar 

  7. Graf J, Yamada Y, Robey FA, Sasaki M, Ogle RC, Kleinman HK, Martin GR (1987) A pentapeptide from the B1 chain of laminin cell adhesion and binds the 67 000 laminin receptor. Biochemistry 26: 6896–6900

    Google Scholar 

  8. Hart IR, Fidler IJ (1978) An in vitro quantitative assay for tumor cell invasion. Cancer Res 38: 3218–3224

    Google Scholar 

  9. Iwamoto Y, Robey FA, Graf J, Sasaki M, Kleinman HK, Yamada Y, Martin GR (1987) YIGSR, a sythetic laminin pentapeptide, inhibits experimental metastasis formation. Science 238: 1132–1134

    Google Scholar 

  10. Iwamoto Y, Reich R, Nemeth G, Yamada Y, Martin GR (1993) Cyclic AMP decreases chemotaxis, invasiveness and lung colonization of H-ras transformed mouse fibroblasts. Clin Expl Metastasis 11: 492–501

    Google Scholar 

  11. Jainckill JL, Aaronson SA, Todaro GJ (1969) Murine sarcoma and leukemia viruses; assay using clonal lines of contact-inhibited mouse cells. J Virol 4: 549–553

    Google Scholar 

  12. Jacobs JP, Jones EM, Baille JP (1970) Characterization of a human diploid cell designated MRC-5. Nature 227: 168–170

    Google Scholar 

  13. Jones PA, DeClerk YA (1980) Destruction of extracellular matrices containing glycoproteins, elastin and collagen by metastatic human tumor cells. Cancer Res 40: 3222–3227

    Google Scholar 

  14. Kleinman HK, MacGarvey ML, Hassel JR, Star VL, Cannon FB, Laurie GW, Martin GR (1986) Basement membrane complexes with biological activity. Biochemistry 25: 312–318

    Google Scholar 

  15. Kramer RH, Vogel KG (1984) Selective degradation of basement membrane macromolecules by metastatic melanoma cells. J Natl Cancer Inst 72: 8889–8899

    Google Scholar 

  16. Laurie GW, Leblond CP, Martin GR (1982) Localization of type IV collagen, laminin, heparansulfate proteoglycan and fibronectin to the basal lamina of basement membranes. J Cell Biol 95: 340–344

    Google Scholar 

  17. Lesot H, Kuhn K, von der Mark K (1985) Isolation of a laminin binding protein from muscle cell membrane. EMBO J 2: 861–865

    Google Scholar 

  18. Levine E, Brik DE, Raska K (1984) Attachment and degradation of collagen substrate by adenovirus-transformed cells of varying tumorigenicity. Collagen Relat Res 4: 49–62

    Google Scholar 

  19. Liotta LA, Lee WC, Morakis DJ (1980) New method for preparing large surfaces of intact basement membranes for tumor invasion studies. Cancer Lett 11: 141–147

    Google Scholar 

  20. Liotta LA, Thorgeirsson UP, Garbisa S (1982) Role of collagenase in tumor cell invasion. Cancer Met Rev 1: 277–288

    Google Scholar 

  21. Liotta LA (1984) Tumor invasion and metastases: role of the basement membrane. Am J Pathol 117: 339–348

    Google Scholar 

  22. MacCarthy JB, Basara ML, Palm SL, Sa DF, Furcht FT (1985) The role of cell adhesion proteins, laminin and fibronectin in the movement of malignant and metastatic cells. Cancer Metastasis Rev 4: 125–152

    Google Scholar 

  23. Malinoff HL, Wicha MS (1983) Isolation of cell surface receptor protein for laminin from murine sarcoma cells. J Cell Biol 96: 1475–1479

    Google Scholar 

  24. Mareel M, Kint J, Meyvisch C (1979) Method of study of malignant C3H fibroblasts into embryonic chick heart in vitro. Virchows Arch B Cell Pathol 30: 95–111

    Google Scholar 

  25. Martin GR, Timpl R (1987) Laminin and other basement membrane components. Ann Rev Biochem 55: 1037–1057

    Google Scholar 

  26. Menzel J, Borth W (1983) Influence of plasma fibronectin on collagenase by collagenase. Collagen Relat Res 3: 217–230

    Google Scholar 

  27. Nicolson GL (1982) Cancer metastasis: organ colonization and the cell surface properties of malignant cells. Biochem Biophys Acta 695: 113–176

    Google Scholar 

  28. Ossowski L, Reich E (1980) Experimental model for quantitative study of metastasis. Cancer Res 40: 2300–2309

    Google Scholar 

  29. Poste G, Doll J, Hart IR, Fidler IJ (1980) In vitro selection of murine B16 melanoma variants with enhanced tissue-invasive properties. Cancer Res 38: 3218–3224

    Google Scholar 

  30. Rao CN, Barsky SH, Terranova VP, Liotta LA (1983) Isolation of a tumor cell laminin receptor. Biochem Biophys Res Commun 111: 804–808

    Google Scholar 

  31. Rasheed S, Nelson-Rees WA, Toth EM, Arnstein P, Gardner MB (1974) Characterization of a newly derived human fibrosarcoma cell line (HT-1080). Cancer 33: 1027–1033

    Google Scholar 

  32. Reddy EP, Reynold RK, Santos E, Barbacid MA (1982) A point mutation is responsible for the aquisition of transforming properties by the T24 bladder carcinoma oncogene. Nature 300: 149–152

    Google Scholar 

  33. Reznikoff CA, Brankow DW, Heidelberger C (1973) Establishment of a clone of C3H mouse embryo cells sensitive to postconfluence inhibition of devision. Cancer Res 33: 3231–3238

    Google Scholar 

  34. Starkey JR, Hosick HL, Stanford DR, Liggit HD (1984) Interaction of metastatic tumor cells with bovine lens. Cancer Res 44: 1585–1594

    Google Scholar 

  35. Taniguchi S, Tatuka S, Nakamatsu K, Inoue M, Sadano H, Okazaki H, Iwamoto Y, Baba T (1989) High invasiveness associated with augumentation in fos-transfered highly metastatic rat 3Y1 cell line. Cancer Res 49: 6738–6744

    Google Scholar 

  36. Tatsuka M, Jinno S, Kakunaga T (1989) Quantitative measurement of cell motility associated with transformed phenotype. Jpn J Cancer Res 80: 408–412

    Google Scholar 

  37. Terranova VP, Liotta LA, Russo RG, Martin GR (1982) Role of laminin in the attachment and metastasis of murine tumor cells. Cancer Res 42: 2265–2269

    Google Scholar 

  38. Thorgeirsson UP, Turpeenniemi-Hujanen T, Williams JE, Westin EH, Heilman CA, Talmadge JE, Liotta LA (1985) NIH3T3 cells transfected with human tumor DNA containing activated ras oncogene express the metastatic phenotype in nude mice. Mol Cell Biol 5: 259–262

    Google Scholar 

  39. Timpl R, Rohde H, Gehron Robey P, Rennard SI, Foidart JM, Martin GR (1979) Laminin-a glycoprotein from basement membrane. J Biol Chem 254: 9933–9937

    Google Scholar 

  40. Timpl R, Fujiwara S, Dziadek M, Aumilley M, Weber S, Engel J (1984) In: Porter, R, Whelan J (eds) Basement membrane and cell movement. Pitman, London, pp 25–74

    Google Scholar 

  41. Timpl R, Dziadek M (1986) Structure, development, and molecular pathology of basement membranes. Int Rev Exp Pathol 29: 1–112

    Google Scholar 

  42. Turpeenniemi-Hujanen T, Thogeirsson UP, Rao CN, Liotta LA (1986) Laminin increases the type IV collagenase from malignant cells. J Biol Chem 261: 1883–1889

    Google Scholar 

  43. Volk T, Geiger B, and Raz B (1984) Motility and adhesive properties of high- and low-metastatic neoplastic cells. Cancer Res 44: 811–824

    Google Scholar 

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Faculty of Medicine, Kyushu University, Maidashi 3-1-1, Higashi-ku, 812, Fukuoka

    Y. Iwamoto, K. Tanaka, K. Okuyama & Y. Sugioka

  2. Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan

    S. Taniguchi

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  1. Y. Iwamoto
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  2. K. Tanaka
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  3. K. Okuyama
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  4. Y. Sugioka
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  5. S. Taniguchi
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Iwamoto, Y., Tanaka, K., Okuyama, K. et al. In vitro assay of the invasive potential of malignant bone and soft tissue tumours through basement membranes. International Orthopaedics 18, 240–247 (1994). https://doi.org/10.1007/BF00188329

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  • DOI: https://doi.org/10.1007/BF00188329

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