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. 2001 Jul;85(1):55–63. doi: 10.1054/bjoc.2001.1854

Differential expression and localization of TIMP-1 and TIMP-4 in human gliomas

L L Groft 1, H Muzik 1, N B Rewcastle 2,6, R N Johnston 1, V Knäuper 3, M A Lafleur 3, P A Forsyth 2,4,5, D R Edwards 3
PMCID: PMC2363922  PMID: 11437402

Abstract

Studies have suggested that an imbalance of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) may contribute to the malignant phenotype of gliomas. In this study, we have undertaken a detailed analysis of expression of the TIMP family in normal human brain and malignant gliomas at both the mRNA and protein level. Reverse transcription-PCR (RT-PCR) analyses of total RNA from surgical tumour specimens revealed unique expression patterns for the 4 members of the TIMP family, with TIMP-1 and -4 showing positive and negative correlations, respectively, with glioma malignancy. By RT-PCR, TIMP-2 and TIMP-3 expression did not change with tumour grade. In situ hybridization localized TIMP-1 to glial tumour cells and also to the surrounding tumour vasculature. TIMP-4 transcripts were predominantly localized to tumour cells, though minor expression was found in vessels. Recombinant TIMP-4 reduced invasion of U251 glioma cells through Matrigel, and U87 clones overexpressing TIMP-4 showed reduced invasive capacity in vitro. TIMP-4, but not TIMP-1, blocked Membrane Type-1-MMP-mediated progelatinase-A (MMP-2) activation in human umbilical vein endothelial cells. The differential expression and localization of individual TIMPs may contribute to the pathophysiology of human malignant gliomas, particularly with regard to tumour vascularization. © 2001 Cancer Research Campaign http://www.bjcancer.com

Keywords: gliomas, metalloproteinases, TIMPs, RT-PCR

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Selected References

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  1. Ahonen M., Baker A. H., Kähäri V. M. Adenovirus-mediated gene delivery of tissue inhibitor of metalloproteinases-3 inhibits invasion and induces apoptosis in melanoma cells. Cancer Res. 1998 Jun 1;58(11):2310–2315. [PubMed] [Google Scholar]
  2. Apte S. S., Olsen B. R., Murphy G. The gene structure of tissue inhibitor of metalloproteinases (TIMP)-3 and its inhibitory activities define the distinct TIMP gene family. J Biol Chem. 1995 Jun 16;270(24):14313–14318. doi: 10.1074/jbc.270.24.14313. [DOI] [PubMed] [Google Scholar]
  3. Bachman K. E., Herman J. G., Corn P. G., Merlo A., Costello J. F., Cavenee W. K., Baylin S. B., Graff J. R. Methylation-associated silencing of the tissue inhibitor of metalloproteinase-3 gene suggest a suppressor role in kidney, brain, and other human cancers. Cancer Res. 1999 Feb 15;59(4):798–802. [PubMed] [Google Scholar]
  4. Baker A. H., Zaltsman A. B., George S. J., Newby A. C. Divergent effects of tissue inhibitor of metalloproteinase-1, -2, or -3 overexpression on rat vascular smooth muscle cell invasion, proliferation, and death in vitro. TIMP-3 promotes apoptosis. J Clin Invest. 1998 Mar 15;101(6):1478–1487. doi: 10.1172/JCI1584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beliën A. T., Paganetti P. A., Schwab M. E. Membrane-type 1 matrix metalloprotease (MT1-MMP) enables invasive migration of glioma cells in central nervous system white matter. J Cell Biol. 1999 Jan 25;144(2):373–384. doi: 10.1083/jcb.144.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bertaux B., Hornebeck W., Eisen A. Z., Dubertret L. Growth stimulation of human keratinocytes by tissue inhibitor of metalloproteinases. J Invest Dermatol. 1991 Oct;97(4):679–685. doi: 10.1111/1523-1747.ep12483956. [DOI] [PubMed] [Google Scholar]
  7. Blavier L., Henriet P., Imren S., Declerck Y. A. Tissue inhibitors of matrix metalloproteinases in cancer. Ann N Y Acad Sci. 1999 Jun 30;878:108–119. doi: 10.1111/j.1749-6632.1999.tb07677.x. [DOI] [PubMed] [Google Scholar]
  8. Butler G. S., Will H., Atkinson S. J., Murphy G. Membrane-type-2 matrix metalloproteinase can initiate the processing of progelatinase A and is regulated by the tissue inhibitors of metalloproteinases. Eur J Biochem. 1997 Mar 1;244(2):653–657. doi: 10.1111/j.1432-1033.1997.t01-1-00653.x. [DOI] [PubMed] [Google Scholar]
  9. Chambers A. F., Matrisian L. M. Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst. 1997 Sep 3;89(17):1260–1270. doi: 10.1093/jnci/89.17.1260. [DOI] [PubMed] [Google Scholar]
  10. DeClerck Y. A., Perez N., Shimada H., Boone T. C., Langley K. E., Taylor S. M. Inhibition of invasion and metastasis in cells transfected with an inhibitor of metalloproteinases. Cancer Res. 1992 Feb 1;52(3):701–708. [PubMed] [Google Scholar]
  11. DeClerck Y. A., Yean T. D., Chan D., Shimada H., Langley K. E. Inhibition of tumor invasion of smooth muscle cell layers by recombinant human metalloproteinase inhibitor. Cancer Res. 1991 Apr 15;51(8):2151–2157. [PubMed] [Google Scholar]
  12. Deryugina E. I., Bourdon M. A., Reisfeld R. A., Strongin A. Remodeling of collagen matrix by human tumor cells requires activation and cell surface association of matrix metalloproteinase-2. Cancer Res. 1998 Aug 15;58(16):3743–3750. [PubMed] [Google Scholar]
  13. Foda H. D., George S., Conner C., Drews M., Tompkins D. C., Zucker S. Activation of human umbilical vein endothelial cell progelatinase A by phorbol myristate acetate: a protein kinase C-dependent mechanism involving a membrane-type matrix metalloproteinase. Lab Invest. 1996 Feb;74(2):538–545. [PubMed] [Google Scholar]
  14. Forsyth P. A., Wong H., Laing T. D., Rewcastle N. B., Morris D. G., Muzik H., Leco K. J., Johnston R. N., Brasher P. M., Sutherland G. Gelatinase-A (MMP-2), gelatinase-B (MMP-9) and membrane type matrix metalloproteinase-1 (MT1-MMP) are involved in different aspects of the pathophysiology of malignant gliomas. Br J Cancer. 1999 Apr;79(11-12):1828–1835. doi: 10.1038/sj.bjc.6990291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gasson J. C., Golde D. W., Kaufman S. E., Westbrook C. A., Hewick R. M., Kaufman R. J., Wong G. G., Temple P. A., Leary A. C., Brown E. L. Molecular characterization and expression of the gene encoding human erythroid-potentiating activity. 1985 Jun 27-Jul 3Nature. 315(6022):768–771. doi: 10.1038/315768a0. [DOI] [PubMed] [Google Scholar]
  16. Greene J., Wang M., Liu Y. E., Raymond L. A., Rosen C., Shi Y. E. Molecular cloning and characterization of human tissue inhibitor of metalloproteinase 4. J Biol Chem. 1996 Nov 29;271(48):30375–30380. doi: 10.1074/jbc.271.48.30375. [DOI] [PubMed] [Google Scholar]
  17. Guedez L., Courtemanch L., Stetler-Stevenson M. Tissue inhibitor of metalloproteinase (TIMP)-1 induces differentiation and an antiapoptotic phenotype in germinal center B cells. Blood. 1998 Aug 15;92(4):1342–1349. [PubMed] [Google Scholar]
  18. Hayakawa T., Yamashita K., Ohuchi E., Shinagawa A. Cell growth-promoting activity of tissue inhibitor of metalloproteinases-2 (TIMP-2). J Cell Sci. 1994 Sep;107(Pt 9):2373–2379. doi: 10.1242/jcs.107.9.2373. [DOI] [PubMed] [Google Scholar]
  19. Hayakawa T., Yamashita K., Tanzawa K., Uchijima E., Iwata K. Growth-promoting activity of tissue inhibitor of metalloproteinases-1 (TIMP-1) for a wide range of cells. A possible new growth factor in serum. FEBS Lett. 1992 Feb 17;298(1):29–32. doi: 10.1016/0014-5793(92)80015-9. [DOI] [PubMed] [Google Scholar]
  20. Hiraoka N., Allen E., Apel I. J., Gyetko M. R., Weiss S. J. Matrix metalloproteinases regulate neovascularization by acting as pericellular fibrinolysins. Cell. 1998 Oct 30;95(3):365–377. doi: 10.1016/s0092-8674(00)81768-7. [DOI] [PubMed] [Google Scholar]
  21. Holten-Andersen M. N., Murphy G., Nielsen H. J., Pedersen A. N., Christensen I. J., Høyer-Hansen G., Brünner N., Stephens R. W. Quantitation of TIMP-1 in plasma of healthy blood donors and patients with advanced cancer. Br J Cancer. 1999 May;80(3-4):495–503. doi: 10.1038/sj.bjc.6690384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kachra Z., Beaulieu E., Delbecchi L., Mousseau N., Berthelet F., Moumdjian R., Del Maestro R., Béliveau R. Expression of matrix metalloproteinases and their inhibitors in human brain tumors. Clin Exp Metastasis. 1999;17(7):555–566. doi: 10.1023/a:1006760632766. [DOI] [PubMed] [Google Scholar]
  23. Khokha R., Waterhouse P., Yagel S., Lala P. K., Overall C. M., Norton G., Denhardt D. T. Antisense RNA-induced reduction in murine TIMP levels confers oncogenicity on Swiss 3T3 cells. Science. 1989 Feb 17;243(4893):947–950. doi: 10.1126/science.2465572. [DOI] [PubMed] [Google Scholar]
  24. Kossakowska A. E., Urbanski S. J., Edwards D. R. Tissue inhibitor of metalloproteinases-1 (TIMP-1) RNA is expressed at elevated levels in malignant non-Hodgkin's lymphomas. Blood. 1991 Jun 1;77(11):2475–2481. [PubMed] [Google Scholar]
  25. Lampert K., Machein U., Machein M. R., Conca W., Peter H. H., Volk B. Expression of matrix metalloproteinases and their tissue inhibitors in human brain tumors. Am J Pathol. 1998 Aug;153(2):429–437. doi: 10.1016/S0002-9440(10)65586-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Leco K. J., Apte S. S., Taniguchi G. T., Hawkes S. P., Khokha R., Schultz G. A., Edwards D. R. Murine tissue inhibitor of metalloproteinases-4 (Timp-4): cDNA isolation and expression in adult mouse tissues. FEBS Lett. 1997 Jan 20;401(2-3):213–217. doi: 10.1016/s0014-5793(96)01474-3. [DOI] [PubMed] [Google Scholar]
  27. Leco K. J., Khokha R., Pavloff N., Hawkes S. P., Edwards D. R. Tissue inhibitor of metalloproteinases-3 (TIMP-3) is an extracellular matrix-associated protein with a distinctive pattern of expression in mouse cells and tissues. J Biol Chem. 1994 Mar 25;269(12):9352–9360. [PubMed] [Google Scholar]
  28. Liotta L. A., Steeg P. S., Stetler-Stevenson W. G. Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell. 1991 Jan 25;64(2):327–336. doi: 10.1016/0092-8674(91)90642-c. [DOI] [PubMed] [Google Scholar]
  29. Llano E., Pendás A. M., Freije J. P., Nakano A., Knäuper V., Murphy G., López-Otin C. Identification and characterization of human MT5-MMP, a new membrane-bound activator of progelatinase a overexpressed in brain tumors. Cancer Res. 1999 Jun 1;59(11):2570–2576. [PubMed] [Google Scholar]
  30. Mohanam S., Wang S. W., Rayford A., Yamamoto M., Sawaya R., Nakajima M., Liotta L. A., Nicolson G. L., Stetler-Stevenson W. G., Rao J. S. Expression of tissue inhibitors of metalloproteinases: negative regulators of human glioblastoma invasion in vivo. Clin Exp Metastasis. 1995 Jan;13(1):57–62. doi: 10.1007/BF00144019. [DOI] [PubMed] [Google Scholar]
  31. Murphy A. N., Unsworth E. J., Stetler-Stevenson W. G. Tissue inhibitor of metalloproteinases-2 inhibits bFGF-induced human microvascular endothelial cell proliferation. J Cell Physiol. 1993 Nov;157(2):351–358. doi: 10.1002/jcp.1041570219. [DOI] [PubMed] [Google Scholar]
  32. Murphy G., Knäuper V. Relating matrix metalloproteinase structure to function: why the "hemopexin" domain? Matrix Biol. 1997 Mar;15(8-9):511–518. doi: 10.1016/s0945-053x(97)90025-1. [DOI] [PubMed] [Google Scholar]
  33. Nagase H., Woessner J. F., Jr Matrix metalloproteinases. J Biol Chem. 1999 Jul 30;274(31):21491–21494. doi: 10.1074/jbc.274.31.21491. [DOI] [PubMed] [Google Scholar]
  34. Nakada M., Nakamura H., Ikeda E., Fujimoto N., Yamashita J., Sato H., Seiki M., Okada Y. Expression and tissue localization of membrane-type 1, 2, and 3 matrix metalloproteinases in human astrocytic tumors. Am J Pathol. 1999 Feb;154(2):417–428. doi: 10.1016/S0002-9440(10)65288-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nakano A., Tani E., Miyazaki K., Yamamoto Y., Furuyama J. Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas. J Neurosurg. 1995 Aug;83(2):298–307. doi: 10.3171/jns.1995.83.2.0298. [DOI] [PubMed] [Google Scholar]
  36. Phillips B. W., Sharma R., Leco P. A., Edwards D. R. A sequence-selective single-strand DNA-binding protein regulates basal transcription of the murine tissue inhibitor of metalloproteinases-1 (Timp-1) gene. J Biol Chem. 1999 Aug 6;274(32):22197–22207. doi: 10.1074/jbc.274.32.22197. [DOI] [PubMed] [Google Scholar]
  37. Powe D. G., Brough J. L., Carter G. I., Bailey E. M., Stetler-Stevenson W. G., Turner D. R., Hewitt R. E. TIMP-3 mRNA expression is regionally increased in moderately and poorly differentiated colorectal adenocarcinoma. Br J Cancer. 1997;75(11):1678–1683. doi: 10.1038/bjc.1997.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Price A., Shi Q., Morris D., Wilcox M. E., Brasher P. M., Rewcastle N. B., Shalinsky D., Zou H., Appelt K., Johnston R. N. Marked inhibition of tumor growth in a malignant glioma tumor model by a novel synthetic matrix metalloproteinase inhibitor AG3340. Clin Cancer Res. 1999 Apr;5(4):845–854. [PubMed] [Google Scholar]
  39. Sawaya R. E., Yamamoto M., Gokaslan Z. L., Wang S. W., Mohanam S., Fuller G. N., McCutcheon I. E., Stetler-Stevenson W. G., Nicolson G. L., Rao J. S. Expression and localization of 72 kDa type IV collagenase (MMP-2) in human malignant gliomas in vivo. Clin Exp Metastasis. 1996 Jan;14(1):35–42. doi: 10.1007/BF00157684. [DOI] [PubMed] [Google Scholar]
  40. Smith M. R., Kung H., Durum S. K., Colburn N. H., Sun Y. TIMP-3 induces cell death by stabilizing TNF-alpha receptors on the surface of human colon carcinoma cells. Cytokine. 1997 Oct;9(10):770–780. doi: 10.1006/cyto.1997.0233. [DOI] [PubMed] [Google Scholar]
  41. Stetler-Stevenson M., Mansoor A., Lim M., Fukushima P., Kehrl J., Marti G., Ptaszynski K., Wang J., Stetler-Stevenson W. G. Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in reactive and neoplastic lymphoid cells. Blood. 1997 Mar 1;89(5):1708–1715. [PubMed] [Google Scholar]
  42. Uhm J. H., Dooley N. P., Villemure J. G., Yong V. W. Glioma invasion in vitro: regulation by matrix metalloprotease-2 and protein kinase C. Clin Exp Metastasis. 1996 Oct;14(5):421–433. doi: 10.1007/BF00128958. [DOI] [PubMed] [Google Scholar]
  43. Uhm J. H., Dooley N. P., Villemure J. G., Yong V. W. Mechanisms of glioma invasion: role of matrix-metalloproteinases. Can J Neurol Sci. 1997 Feb;24(1):3–15. doi: 10.1017/s0317167100021028. [DOI] [PubMed] [Google Scholar]
  44. Velasco G., Cal S., Merlos-Suárez A., Ferrando A. A., Alvarez S., Nakano A., Arribas J., López-Otín C. Human MT6-matrix metalloproteinase: identification, progelatinase A activation, and expression in brain tumors. Cancer Res. 2000 Feb 15;60(4):877–882. [PubMed] [Google Scholar]
  45. Wang M., Liu Y. E., Greene J., Sheng S., Fuchs A., Rosen E. M., Shi Y. E. Inhibition of tumor growth and metastasis of human breast cancer cells transfected with tissue inhibitor of metalloproteinase 4. Oncogene. 1997 Jun 12;14(23):2767–2774. doi: 10.1038/sj.onc.1201245. [DOI] [PubMed] [Google Scholar]
  46. Will H., Atkinson S. J., Butler G. S., Smith B., Murphy G. The soluble catalytic domain of membrane type 1 matrix metalloproteinase cleaves the propeptide of progelatinase A and initiates autoproteolytic activation. Regulation by TIMP-2 and TIMP-3. J Biol Chem. 1996 Jul 19;271(29):17119–17123. doi: 10.1074/jbc.271.29.17119. [DOI] [PubMed] [Google Scholar]
  47. Wingfield P. T., Sax J. K., Stahl S. J., Kaufman J., Palmer I., Chung V., Corcoran M. L., Kleiner D. E., Stetler-Stevenson W. G. Biophysical and functional characterization of full-length, recombinant human tissue inhibitor of metalloproteinases-2 (TIMP-2) produced in Escherichia coli. Comparison of wild type and amino-terminal alanine appended variant with implications for the mechanism of TIMP functions. J Biol Chem. 1999 Jul 23;274(30):21362–21368. doi: 10.1074/jbc.274.30.21362. [DOI] [PubMed] [Google Scholar]
  48. Wong H., Anderson W. D., Cheng T., Riabowol K. T. Monitoring mRNA expression by polymerase chain reaction: the "primer-dropping" method. Anal Biochem. 1994 Dec;223(2):251–258. doi: 10.1006/abio.1994.1581. [DOI] [PubMed] [Google Scholar]
  49. Yamamoto M., Mohanam S., Sawaya R., Fuller G. N., Seiki M., Sato H., Gokaslan Z. L., Liotta L. A., Nicolson G. L., Rao J. S. 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. 1996 Jan 15;56(2):384–392. [PubMed] [Google Scholar]
  50. Yong V. W., Krekoski C. A., Forsyth P. A., Bell R., Edwards D. R. Matrix metalloproteinases and diseases of the CNS. Trends Neurosci. 1998 Feb;21(2):75–80. doi: 10.1016/s0166-2236(97)01169-7. [DOI] [PubMed] [Google Scholar]
  51. Yoshiji H., Harris S. R., Raso E., Gomez D. E., Lindsay C. K., Shibuya M., Sinha C. C., Thorgeirsson U. P. Mammary carcinoma cells over-expressing tissue inhibitor of metalloproteinases-1 show enhanced vascular endothelial growth factor expression. Int J Cancer. 1998 Jan 5;75(1):81–87. doi: 10.1002/(sici)1097-0215(19980105)75:1<81::aid-ijc13>3.0.co;2-g. [DOI] [PubMed] [Google Scholar]
  52. Zeng Z. S., Cohen A. M., Zhang Z. F., Stetler-Stevenson W., Guillem J. G. Elevated tissue inhibitor of metalloproteinase 1 RNA in colorectal cancer stroma correlates with lymph node and distant metastases. Clin Cancer Res. 1995 Aug;1(8):899–906. [PubMed] [Google Scholar]
  53. Zhou Z., Apte S. S., Soininen R., Cao R., Baaklini G. Y., Rauser R. W., Wang J., Cao Y., Tryggvason K. Impaired endochondral ossification and angiogenesis in mice deficient in membrane-type matrix metalloproteinase I. Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4052–4057. doi: 10.1073/pnas.060037197. [DOI] [PMC free article] [PubMed] [Google Scholar]

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