Skip to main content

Advertisement

Log in

The Expression and Activation of Matrix Metalloproteinase-2 in Rat Brain After Implantation of C6 Rat Glioma Cells

  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

It has been reported that matrix metalloproteinases (MMPs) are highly expressed in malignant glioma cells and that this increased expression may facilitate the invasiveness of tumor cells. The authors investigated the expression and enzymatic activity of MMPs in rat brain during the growth of malignant gliomas at different time intervals. C6 rat glioma cells were unilaterally implanted into rat cerebral hemispheres. After 7 or 14 days, these brain tissues were prepared for SDS-PAGE zymography, Western blotting, immunohistochemistry, and in situ zymography. SDS-PAGE zymography and Western blotting revealed that the expression of proMMP-2 in rat brains with C6 glioma cells was significantly higher than that in normal or the sham-operated rat brains, and that the activated form of MMP-2 was detected only in the former but not in the latter. On immunohistochemistry, C6 glioma cells presenting invasive growth into the rat brain parenchyma and vessels demonstrated MMP-2 immunoreactivity. On in situ zymography, foci of invasive C6 glioma cells in rat brain tissue showed gelatinolytic activity. These results suggest that expression and activation of MMP-2 may be one of the crucial steps for glioma cell invasion into the brain parenchyma in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Liotta LA, Steeg PS, Stetler-Stevenson WG: Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell 64: 327-336, 1991

    Google Scholar 

  2. Apodaca G, Rutka JT, Bouhana K, Berens ME, Giblin JR, Rosenblum ML, McKerrow JH, Banda MJ: Expression of metalloproteinases and metalloproteinase inhibitors by fetal astrocytes and glioma cells. Cancer Res 50: 2322-2329, 1990

    Google Scholar 

  3. Nakagawa T, Kubota T, Kabuto M, Sato K, Kawano H, Hayakawa T, Okada Y: Production of matrix metalloproteinases and tissue inhibitor of metalloproteinase-1 by human brain tumors. J Neurosurg 81: 69-77, 1994

    Google Scholar 

  4. Nakano A, Tani E, Miyazaki K, Yamamoto Y, Furuyama J: Matrix metalloproteinases and tissue inhibitor of metalloproteinases in human gliomas. J Neurosurg 83: 298-307, 1995

    Google Scholar 

  5. Sawaya RE, Yamamoto M, Gokaslan ZL, Wang SW, Mohanam S, Fuller GN, McCutcheon IE, Stetler-Stevenson WG, Nicolson GL, Rao JS: Expression and localization of 72 kDa type IV collagenase (MMP-2) in human malignant gliomas in vivo. Clin Exp Metastasis 14: 35-42, 1996

    Google Scholar 

  6. Abe T, Mori T, Kohno K, Seiki M, Hayakawa T, Welgus HG, Hori S, Kuwano M: Expression of 72 kD type IV collagenase and invasion activity of human glioma cells. Clin Exp Metastasis 12: 296-304, 1994

    Google Scholar 

  7. Chintala SK, Sawaya R, Gokaslan ZL, Rao JS: Modulation of matrix metalloprotease-2 and invasion in human glioma cells by α3β1 integrin. Cancer Lett 103: 201-208, 1996

    Google Scholar 

  8. Deryugina EI, Luo GX, Reisfeld RA, Bourdon MA, Strongin A: Tumor cell invasion through Matrigel is regulated by activated matrix metalloproteinase-2. Anticancer Res 17: 3201-3210, 1997

    Google Scholar 

  9. Nakagawa T, Kubota T, Kabuto M, Kodera T: Captopril inhibits glioma invasion in vitro: involvement of matrix metalloproteinases. Anticancer Res 15: 1985-1991, 1995

    Google Scholar 

  10. Nakagawa T, Kubota T, Kabuto M, Fujimoto N, Okada Y: Secretion of matrix metalloproteinase-2 (72 kD gelatinase/type IV collagenase D gelatinase A) by malignant human glioma cell lines: implications for the growth and cellular invasion of the extracellular matrix. J Neuro-Oncol 28: 13-24, 1996

    Google Scholar 

  11. Rutka JT, Matsuzawa K, Hubbard SL, Furuyama K, Becker LE, Stetler-Stevenson W, Edwards DR, Dirks PB: 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: 877-884, 1995

    Google Scholar 

  12. Uhm HJ, Dooley NP, Villemure J-G, Yong VW: Glima invasion in vitro: regulation by matrix metalloprotease-2 and protein kinase C. Clin Exp Metast 14: 421-433, 1996

    Google Scholar 

  13. Sawaya R, Go Y, Kyritisis AP, Uhm J, Venkaiah B, Mohanam S, Gokaslan ZL, Rao JS: Elevated levels of Mr 92,000 type IV collagenase during tumor growth in vivo. Biochem Biophys Res Commun 251: 632-636, 1998

    Google Scholar 

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

    Google Scholar 

  15. Hibbs MS, Hasty KA, Seyer JM, Kang AH, Mainardi CL: Biochemical and immunological characterization of the secreted forms of human neutrophil gelatinase. J Biol Chem 260: 2493-2500, 1985

    Google Scholar 

  16. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci USA 76: 4350-4354, 1979

    Google Scholar 

  17. Galis ZS, Sukhova GK, Lark MW, Libby P: Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques. J Clin Invest 94: 2493-2503, 1994

    Google Scholar 

  18. Galis ZS, Sukhova GK, Libby P: Microscopic localization of active proteases by in situ zymography: detection of matrix metalloproteinase activity in vascular tissue. FASEB J 9: 974-980, 1995

    Google Scholar 

  19. Tamaki M, McDonald W, Amberger VR, Moore E, Del Maestro RF: Implantation of C6 astrocytoma spheroid into collagen type I gels: invasive, proliferative, and enzymatic characterizations. J Neurosurg 87: 602-609, 1997

    Google Scholar 

  20. de Ridder L, Calliauw L: Invasiveness of primary and secondary brain tumors in vitro correlated with clinical results. Neurosurgery 31: 1043-1048, 1992

    Google Scholar 

  21. Paganetti PA, Caroni P, Schwab ME: Glioblastoma infiltration into central nervous system tissue in vitro: involvement of a metalloprotease. J Cell Biol 107: 2281-2291, 1988

    Google Scholar 

  22. Lund-Johansen M, Rucklidge GJ, Milne G, Bjerkvig R: A metalloproteinase, capable of destroying cultured brain tissue isolated from rat glioma cells. Anticancer Res 11: 1001-1006, 1991

    Google Scholar 

  23. Benda P, Someda K, Messer J, Sweet WH: Morphological and immunochemical studies of rat glial tumors and clonal strains propagated in culture. J Neurosurg 34: 310-323, 1971

    Google Scholar 

  24. Barth RF: Rat brain tumor models in experimental neurooncology: The 9L, C6, T9, F98, RG2 (D74), RT-2 and CNS-1 gliomas. J Neuro-Oncol 36: 91-102, 1998

    Google Scholar 

  25. Chicoine MR, Silbergeld DL: Invading C6 glioma cells maintaining tumorigenicity. J Neurosurg 83: 665-671, 1995

    Google Scholar 

  26. Nagano N, Sasaki H, Aoyagi M, Hirakawa K: Invasion of experimental rat brain tumor: early morphological changes following microinjection of C6 glioma cells. Acta Neuropathol 86: 117-125, 1993

    Google Scholar 

  27. Vaithilingam IS, Stroude EC, McDonald W, Del Maestro RF: General protease and collagenase (IV) activity in C6 astrocytoma cells, C6 spheroids and implanted C6 spheroids. J Neuro-Oncol 10: 203-212, 1991

    Google Scholar 

  28. Amberger VR, Paganetti PA, Seulberger H, Eldering JA, Schwab ME: Characterization of a membrane-bound metalloendoprotease of rat C6 glioblastoma cells. Cancer Res 54: 4017-4025, 1994

    Google Scholar 

  29. Del Maestro RF, Vaithilingam IS, McDonald W: Degradation of collagen type IV by C6 astrocytoma cells. J Neuro-Oncol 24: 75-81, 1995

    Google Scholar 

  30. Tamaki M, McDonald W, Del Maestro RF: Release of collagen type IV degrading activity from C6 astrocytoma cells and cell density. J Neurosurg 84: 1013-1019, 1996

    Google Scholar 

  31. Vaithilingam IS, McDonald W, Malott DW, Del Mestro RF: An extracellular proteasome-like structure from C6 astrocytoma cells with serine collagenase IV activity and metallodependent activity on a-casein and a-insulin. J Biol Chem 270: 4588-4593, 1995

    Google Scholar 

  32. Pei D, Weiss SJ: Transmembrane-deletion mutants of the membrane-type matrix metalloproteinase-1 process progelatinase A and express intrinsic matrix-degrading activity. J Biol Chem 271: 9135-9140, 1996

    Google Scholar 

  33. Sato H, Takino T, Okada Y, Cao J, Shinagawa A, Yamamoto E, Seiki M: A matrix metalloproteinase expressed on the surface of invasive tumour cells. Nature 370: 61-65, 1994

    Google Scholar 

  34. Takino T, Sato H, Shinagawa A, Seiki M: Identification of the second membrane-type matrix metalloproteinase (MT-MMP-2) gene from a human placenta cDNA library. J Biol Chem 270: 23013-23020, 1995

    Google Scholar 

  35. Deryugina EI, Bourdon MA, Luo GX, Reisfeld RA, Strongin A: Matrix metalloproteinase-2 activation modulates glioma cell migration. J Cell Sci 110: 2473-2482, 1997

    Google Scholar 

  36. Yamamoto M, Mohanam S, Sawaya R, Fuller GN, Seiki M, Sato H, Gokaslan ZL, Liotta LA, Nicolson GL, Rao JS: Differential expression of membrane-type matrix metalloproteinase and its correlation with gelatinase A activation in human malignant brain tumors in vivo and in vitro. Cancer Res 56: 384-392, 1996

    Google Scholar 

  37. Amberger VR, Hensel T, Ogata N, Schwab ME: Spreading and migration of human glioma and rat C6 cells on central nervous system myelin in vitro is correlated with tumor malignancy and involves a metalloproteolytic activity. Cancer Res 58: 149-158, 1998

    Google Scholar 

  38. Rutka JT, Apodaca G, Stern R, Rosenblum M: The extracellular matrix of the central and peripheral nervous systems: structure and function. J Neurosurg 69: 155-170, 1988

    Google Scholar 

  39. Woessner JF Jr: Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 5: 2145-2154, 1991

    Google Scholar 

  40. Zuo J, Ferguson TA, Hernandez YJ, Stetler-Stevenson WG, Muir D: Neuronal matrix metalloproteinase-2 degrades and inactivates a neurite-inhibiting chondroitin sulfate proteoglycan. J Neurosci 18: 5203-5211, 1998

    Google Scholar 

  41. Knauper V, Cowell S, Smith B, Lopez-Otin C, O'Shea M, Morris H, Zardi L, Murphy G: The role of the C-terminal domain of human collagenase-3 (MMP-13) in the activation of procollagenase-3, substrate specificity, and tissue inhibitor of metalloproteinase interaction. J Biol Chem 272: 7608-7616, 1997

    Google Scholar 

  42. Nakagawa T, Kubota T, Kabuto M, Kodera T: Hyaluronic acid facilitates glioma cell invasion in vitro. Anticancer Res 16: 2917-2922, 1996

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kodera, T., Nakagawa, T., Kubota, T. et al. The Expression and Activation of Matrix Metalloproteinase-2 in Rat Brain After Implantation of C6 Rat Glioma Cells. J Neurooncol 46, 105–114 (2000). https://doi.org/10.1023/A:1006387600909

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006387600909

Navigation