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Glioma invasionin vitro: regulation by matrix metalloprotease-2 and protein kinase C

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A hallmark of invasive tumors is their ability to effect degradation of the surrounding extracellular matrix (ECM) by the local production of proteolytic enzymes, such as the matrix metalloproteases (MMPs). In this paper, we demonstrate that the invasion of human gliomas is mediated by a 72 kDa MMP, referred to as MMP-2, and provide further evidence that the activity of MMP-2 is regulated by protein kinase C (PKC). The invasiveness of five human glioma cell lines (A172, U87, U118, U251, U563) was assessed in anin vitro invasion assay and was found to correlate with the level of MMP-2 activity (r 2 = 0.95); in contrast, the activity of this 72 kDa metalloprotease was barely detectable in non-invasive control glial cells (non-transformed human astrocytes and oligodendrocytes). Treatment with 1,10-phenanthroline, a metalloprotease inhibitor, or with a synthetic dipeptide, containing a blocking sequence (ala-phe) specific for MMPs, resulted in a > 90% reduction in glioma invasion. Furthermore, this MMP-2 activity could be inhibited by the treatment of tumor cells with calphostin C, a specific inhibitor of PKC. Glioma cell lines treated with calphostin C demonstrated a dose-dependent decrease (IC50 = 30 nm) in tumor invasiveness with a concomitant reduction in the activity of the MMP-2. Conversely, treatment of non-invasive control astrocytes with a PKC activator (phorbol ester) led to a corresponding increase in their invasiveness and metalloprotease activity. These findings support the postulate that MMP-2 activity constitutes an important effector of human glioma invasion and that the regulation of this proteolytic activity can be modulated by PKC.

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References

  1. Berens ME, Rutka JT and Rosenblum ML, 1990, Brain tumor epidemiology, growth, and invasion.Neurosurg Clinics N Am,1, 1–18.

    Google Scholar 

  2. Couldwell WT, de Tribolet N, Antel JP, Gauthier T and Kuppner MC, 1992, Adhesion molecules and malignant gliomas: implications for tumorigenesis.J Neurosurg,76, 782–91.

    Google Scholar 

  3. Mignatti P and Rifkin DB, 1993, Biology and biochemistry of proteinases in tumor invasion.Physiol Rev,73, 161–95.

    Google Scholar 

  4. Moscatelli D and Rifkin DB, 1988, Membrane and matrix localization of proteinases: a common theme in tumor cell invasion and angiogenesis.Biochim Biophys Acta,948, 67–85.

    Google Scholar 

  5. Vassalli J-D and Pepper MS, 1994, Membrane proteases in focus.Nature,370, 14–15.

    Google Scholar 

  6. Cockett MI, Birch ML, Murphy G, Hart IR and Docherty AJ, 1994, Metalloproteinase domain structure, cellular invasion and metastasis.Biochem Soc Trans,22, 55–7.

    Google Scholar 

  7. Sato H, Takino T, Okada Y,et al. 1994, A matrix metalloproteinase expressed on the surface of invasive tumour cells.Nature,370, 61–5.

    Google Scholar 

  8. Rucklidge GJ, Edvardsen G and Bock E, 1994, Cell-adhesion molecules and metalloproteinases: a linked role in tumour cell invasiveness.Biochem Soc Trans,22,63–8.

    Google Scholar 

  9. Moser TL, Young TN, Rodriguez GC, Pizzo SV, Bast RCJ and Stack MS, 1994, Secretion of extracellular matrix-degrading proteinases is increased in epithelial ovarian carcinoma.Int J Cancer,56, 552–9.

    Google Scholar 

  10. Seftor RE, Seftor EA, Stetler-Stevenson WG and Hendrix MJ, 1993, The 72 kDa type IV collagenase is modulated via differential expression of αvβ3 and α5β1 integrins during human melanoma cell invasion.Cancer Res,53, 3411–15.

    Google Scholar 

  11. Agarwal C, Hembree JR, Rorke EA and Eckert RL, 1994, Transforming growth factor beta 1 regulation of metalloproteinase production in cultured human cervical epithelial cells.Cancer Res,54, 943–9.

    Google Scholar 

  12. Albini A, Melchiori A, Santi L, Liotta LA, Brown PD and Stetler-Stevenson WG, 1991, Tumor cell invasion inhibited by TIMP-2 [see comments].J Natl Cancer Inst,83, 775–9.

    Google Scholar 

  13. Davies B, Waxman J, Wasan H,et al. 1993, Levels of matrix metalloproteases in bladder cancer correlate with tumor grade and invasion.Cancer Res,53, 5365–9.

    Google Scholar 

  14. Lund-Johansen M, Rucklidge GJ, Milne G and Bjerkvig R, 1991, A metalloproteinase, capable of destroying cultured brain tissue isolated from rat glioma cells.Anticancer Res,11, 1001–6.

    Google Scholar 

  15. Vaithilingam IS, McDonald W, Brown NK, Stroude E, Cook RA and Del Maestro RF, 1992, Serum proteolytic activity during the growth of C6 astrocytoma.J Neurosurg,77, 595–600.

    Google Scholar 

  16. Nakagawa T, Kubota T, Kabuto M et al, 1994, Production of matrix metalloproteinases and tissue inhibitor of metalloproteinases-1 by human brain tumors.J Neurosurg,00, 000–000.

    Google Scholar 

  17. Del Maestro RF, Lopez-Torres M, McDonald WB,et al. 1992, The effect of tumor necrosis factor-α on human malignant glial cells.J Neurosurg,76, 652–9.

    Google Scholar 

  18. Paganetti PA, Caroni P and Schwab ME, 1988, Glioblastoma infiltration into central nervous system tissuein vitro: involvement of a metalloprotease.J Cell Biol,107, 2281–91.

    Google Scholar 

  19. Bjerkvig R, Laerum OD and Mella O, 1986, Glioma cell interactions with fetal rat brain aggregatesin vitro and with brain tissuein vivo.Cancer Res,46, 4071–9.

    Google Scholar 

  20. Apodaca G, Rutka JT, Bouhana K,et al. 1990, Expression of metalloproteinases and metalloproteinase inhibitors by fetal astrocytes and glioma cells.Cancer Res,50, 2322–9.

    Google Scholar 

  21. Abe T, Mori T, Kohnoet al. 1994, Expression of 72 kDa type IV collagenase and invasion activity of human glioma cells.Clin Exp Metastasis,12, 296–304.

    Google Scholar 

  22. Couldwell WT, Antel JP, Apuzzo MLJ and Yong VW, 1990, Inhibition of growth of established human glioma lines by modulators of the protein kinase C second messenger system.72, 349. (Abstract).

    Google Scholar 

  23. Couldwell WT, Antel JP, Apuzzo MLJ and Yong VW, 1990, Inhibition of growth of established human glioma lines by modulators of the protein kinase C system.J Neurosurg,73, 594–600.

    Google Scholar 

  24. Couldwell WT, Uhm JH, Antel JP and Yong VW, 1991, Enhanced protein kinase C activity correlates with the growth rate of malignant gliomasin vitro.Neurosurgery,29, 880–7.

    Google Scholar 

  25. Couldwell WT, Antel JP and Yong VW, 1992, Protein kinase C (PKC) activity correlates with growth rate of gliomas. II: Effects of glioma mitogens and modulators of PKC.Neurosurgery,31, 717–24.

    Google Scholar 

  26. Collins VP, 1983, Cultured human glial and glioma cells.Int Rev Exp Pathol,24, 135–92.

    Google Scholar 

  27. Yong VW, Tejada-Berges T, Goodyer CG, Antel JP and Yong FP, 1992, Differential proliferative responses of human and mouse astrocytes to gamma-interferon.Glia,6, 269–80.

    Google Scholar 

  28. Bruns RF, Miller FD, Merriman RL et al. 1991, Inhibition of protein kinase C by calphostin C is lightdependent.Biochem Biophys Res Commun,176, 288–93.

    Google Scholar 

  29. Albini A, Iwamoto Y, Kleinman HKet al. 1987, A rapidin vitro assay for quantitating the invasive potential of tumor cells.Cancer Res,47, 3329–245.

    Google Scholar 

  30. Overall C, Wrana J and Sodek J, 1989, Independent regulation of collagenase, 72 kDa progelatinase and metalloendoproteinase inhibitor expression in human fibroblasts by transforming growth factor-beta.J Biol Chem,264, 1860–9.

    Google Scholar 

  31. Mainardi C, Hasty K and Hibbs M, 1984, Antibody to rabbit macrophage type V collagenase/gelatinase and its use to further characterize the enzyme.Collagen Rel Res,4, 209–17.

    Google Scholar 

  32. Baltuch GH, Couldwell WT, Villemure J-G and Yong VW, 1993, Protein kinase C inhibitors suppress cell growth in established and low passage glioma cell lines. A comparison between staurosporine and tamoxifen.Neurosurgery,33, 495–501.

    Google Scholar 

  33. Ramos-DeSimone N and French DL, 1994, Monoclonal antibodies to human MMP-9.732, 469–71 (Abstract).

    Google Scholar 

  34. Baxter DA, Johnston D and Strittmatter WJ, 1983, Protease inhibitors implicate metalloprotease in synaptic transmission at the mammalian neuromuscular junction.Proc Natl Acad Sci,80, 4174–8.

    Google Scholar 

  35. Stoppelli MP, Verde P, Grimaldi G, Locatelli EK and Blasi F, 1986, Increase in urokinase plasminogen activator mRNA synthesis in human carcinoma cells is a primary effect of the potent tumor promoter, phorbol myristate acetate.J Cell Biol,102, 1235–41.

    Google Scholar 

  36. Basu A, 1993, The potential of protein kinase C as a target for anticancer treatment.Pharmac Ther,59, 257–80.

    Google Scholar 

  37. Schwartz GK, Redwood SM, Ohnuma T, Holland JF, Droller MJ and Liu BC-S, 1990, Inhibition of invasion of invasive human bladder carcinoma cells by protein kinase C inhibitor staurosporine.J Natl Cancer Inst,82, 1753–56.

    Google Scholar 

  38. Matrisian LM, 1994, Matrix metalloproteinase gene expression.A NY Acad Sci,732, 42–50.

    Google Scholar 

  39. Levy AT, Cioce V, Sobel ME, Garbisa S, Grigioni WF and Liotta LA, 1991, Increased expression of theM r 72,000 type IV collagenase in human colonic adenocarcinoma.Cancer Res,51, 439–44.

    Google Scholar 

  40. Liotta LA, Rao NC and Wewer UM, 1986, Biochemical interactions of tumor cells with the basement membrane.A Rev Biochem,55, 1037–57.

    Google Scholar 

  41. Yamagata S, Tanaka R, Ito Y and Shimizu S, 1989, Gelatinases of murine metastatic tumor cells.Biochem Biophys Res Commun,158, 228–34.

    Google Scholar 

  42. Rao JS, Steck PA, Mohanam S, Stetler-Stevenson WG and Liotta LA, 1993, Elevated levels ofM r 92,000 type IV collagenase in human brain tumors.Cancer Res,53, 2208–11.

    Google Scholar 

  43. Rao JS, Steck PA, Tofilonet al. 1994, Role of plasminogen activator and of 92-kDa type IV collagenase in glioblastoma invasion using anin vitro Matrigel model.J Neurooncol,18, 129–38.

    Google Scholar 

  44. Parisi JE and Scheithauer BW, 1993, Glial tumors. In: Nelson JS, Parisi JE and Schochet SS Jr eds. Principles and Practice of Neuropathology, St Louis: Mosby, pp. 123–84.

    Google Scholar 

  45. Herron GS, Banda MJ, Clark EJ, Gavrilovic J and Werb Z, 1986, Secretion of metalloproteinases by stimulated capillary endothelial cells. II. Expression of collagenase and stromelysin activities is regulated by endogenous inhibitors.J Biol Chem,261, 2814–18.

    Google Scholar 

  46. Herron GS, Werb Z, Dwyer K and Banda MJ, 1986, Secretion of metalloproteinases by stimulated capillary endothelial cells. I. Production of procollagenase and prostromelysin exceeds expression of proteolytic activity.J Biol Chem,261, 2810–13.

    Google Scholar 

  47. Kapila S, Lee C and Richards DW, 1995, Characterization and identification of proteinases and proteinase inhibitors synthesized by temporo-mandibular joint disc cells.J Dent Res,74, 1328–36.

    Google Scholar 

  48. Cook TF, Burke JS, Bergman KD, Quinn CO, Jeffrey JJ and Partridge NC, 1994, Cloning and regulation of rat tissue inhibitor of metalloproteinase-2 in oesteoblastic cells.Arch Biochem Biophys,311, 313–20.

    Google Scholar 

  49. Campbell EJ, Cury JD, Shapiro SD, Goldberg GI and Welgus HG, 1991, Neutral proteinases of human mononuclear phagocytes. Cellular differentiation markedly alters cell phenotype for serine proteinases, metalloproteinases, and tissue inhibitor of metalloproteinases.J Immunol,146, 1286–93.

    Google Scholar 

  50. Fini ME and Girard MT, 1990, The pattern of metalloproteinase expression of corneal fibroblasts is altered by passage in cell culture.J Cell Sci,97, 373–83.

    Google Scholar 

  51. Sottile J, Mann DM, Diemer V and Millis AJ, 1989, Regulation of collagenase and collagenase mRNA production in early- and late-passage human diploid fibroblasts.J Cell Physiol,138, 281–90.

    Google Scholar 

  52. Lamph WW, Wamsley P, Sassone-Corsi P and Verma IM, 1988, Induction of proto-oncogenejun/AP-1 by scrum and TPA.Nature,334, 629–31.

    Google Scholar 

  53. Greenberg ME and Ziff EB, 1984, Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene.Nature,311, 433–8.

    Google Scholar 

  54. Mackay AR, Ballin M, Pelina MD,et al. 1992, Effect of phorbol ester and cytokines of matrix metalloproteinase and tissue inhibitor of metalloproteinase expression in tumor and normal cell lines.Invasion Metast,12, 168–184.

    Google Scholar 

  55. Gopalakrishna R and Barsky SH, 1988, Tumor promoter-induced membrane-bound kinase C regulates hematogenous metastasis.Proc Natl Acad Sci,85, 612–16.

    Google Scholar 

  56. Takenaga K and Takahashi K, 1986, Effects of 12-O-tetradecanylphorbol-13-acetate on adhesiveness and lung-colonizing ability of Lewis lung carcinoma cells.Cancer Res,40, 375–80.

    Google Scholar 

  57. Korczak B, Whale C and Kerbel RS, 1989, Possible involvement of Ca2+ mobilization and protein kinase C activation in the induction of spontaneous metastasis by mouse mammary adenocarcinoma cells.Cancer Res,49, 2597–602.

    Google Scholar 

  58. Nabeshima K, Komada N, Kishi J, Koita H, Inoue T and Hayakawa T, 1993, TPA-enhanced invasion of Matrigel associated with augmentation of cell motility but no metalloproteinase activity in a highly metastasic variant (L-10) of human rectal adenocarcinoma cell line RCM-1.Int J Cancer,55, 974–81.

    Google Scholar 

  59. Schwartz GK, Jiang J, Kelsen D and Albino AP, 1994, Protein kinase C: a novel target for inhibiting gastric cancer cell invasion.J Natl Cancer Inst,85, 402–7.

    Google Scholar 

  60. Gross JL, Moscatelli D, Jaffe EA and Rifkin DB, 1982, Plasminogen activator and collagenase production by cultured capillary endothelial cells.J Cell Biol,95, 974–81.

    Google Scholar 

  61. Overall CM, 1994, Regulation of tissue inhibitor of matrix metalloproteinase expression. In: Greenwald RA and Golub LM, eds.Inhibition of Matrix Metalloproteinases: therapeutic potential. New York: The New York Academy of Sciences, pp. 51–64.

    Google Scholar 

  62. Tanaka K, Iwamoto Y, Ito Y,et al. 1995, Cyclic AMP-regulated synthesis of the tissue inhibitors of metalloproteinases suppresses the invasive potential of the human fibrosarcoma cell line HT1080.Cancer Res,55, 2927–35.

    Google Scholar 

  63. Rutka JT, Matsuzawa K, Hubbard SL,et al. 1995, Expression of TIMP-1, TIMP-2, 72- and 92-kDa type IV collagenase transcripts in human astrocytoma cell lines: correlation with astrocytoma cell invasiveness.Int J Oncol,6, 877–84.

    Google Scholar 

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Authors and Affiliations

  1. Brain Tumor Research Group, Montreal Neurological Institute, Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, Canada

    Joon H. Uhm, Nora P. Dooley, Jean-Guy Villemure & Voon Wee Yong

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  1. Joon H. Uhm
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  2. Nora P. Dooley
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  3. Jean-Guy Villemure
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  4. Voon Wee Yong
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Joon Uhm and Nora Dooley contributed equally to this work.

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Uhm, J.H., Dooley, N.P., Villemure, JG. et al. Glioma invasionin vitro: regulation by matrix metalloprotease-2 and protein kinase C. Clin Exp Metast 14, 421–433 (1996). https://doi.org/10.1007/BF00128958

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