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. 2007 May 1;10(2):499–510. doi: 10.1111/j.1582-4934.2006.tb00415.x

MMP-9 and MMP-2 gelatinases and TIMP-1 and TIMP-2 inhibitors in breast cancer: correlations with prognostic factors

D C Jinga b, A Blidaru b, Ileana Condrea b, Carmen Ardeleanu c, Cristina Dragomir a, Geza Szegli a, Maria Stefanescu a, Cristiana Matache a,*
PMCID: PMC3933137  PMID: 16796815

Abstract

The goal of our study was to analyse the prognostic values for some matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in breast cancer. We evaluated the activity and the expression levels of MMP-9, MMP-2, TIMP-1 and TIMP-2 in malignant versus benign fresh breast tumor extracts. For this purpose, gelatinzymography, immunoblotting and ELISA were used to analyse the activity and expression of MMPs and TIMPs. We found that MMP-9 expression level and activity are increased in malignant tumors. In addition, MMP-9/TIMP-1 and MMP-2/TIMP-2 ratio values obtained by us were significantly different in malignant tumors compared to benign tumors. We suggest that the abnormal MMP-9/TIMP-1 balance plays a role in the configuration of breast invasive carcinoma of no special type and also in tumor growth, while altered MMP-2/TIMP-2 ratio value could be associated with lymph node invasion and used as a prognostic marker in correlation with Nottingham Prognostic Index. Finally, we showed that in malignant tumors high expression of estrogen receptors is associated with enhanced activity of MMP-2 and increased bcl-2 levels, while high expression of progesterone receptors is correlated with low TIMP-1 protein levels.

Keywords: breast cancer, MMP-9, MMP-2, TIMP-1, TIMP-2, clinicopathological markers

References

  • 1.Nagase H, Woessner JF. Matrix metalloproteinases. J Biol Chem. 1999;274:21491–4. doi: 10.1074/jbc.274.31.21491. [DOI] [PubMed] [Google Scholar]
  • 2.Matrisian LM. Metalloproteinases and their inhibitors in matrix remodelling. Trends Genet. 1990;6:121–5. doi: 10.1016/0168-9525(90)90126-q. [DOI] [PubMed] [Google Scholar]
  • 3.Nagase H. Activation mechanisms of matrix metalloproteinases. Biol Chem. 1997;378:151–60. [PubMed] [Google Scholar]
  • 4.Massova I, Kotra LP, Fridman R, Mobashery S. Matrix metalloproteinases: structures, evolution, and diversification. FASEB J. 1998;12:1075–95. [PubMed] [Google Scholar]
  • 5.Benz CC. Transcriptional factors and breast cancer. Endoc Rel Cancer. 1998;5:271–82. [Google Scholar]
  • 6.Giunciuglio D, Culty M, Fassina G, Masiello L, Melchiori A, Paglialunga G, Arand G, Ciardiello F, Basolo F, Thomson EW. Invasive phenotype of MCF10A cells overexpressing c-Ha-ras and c-erbB-2 oncogenes. Int J Cancer. 1995;63:815–22. doi: 10.1002/ijc.2910630612. [DOI] [PubMed] [Google Scholar]
  • 7.George SJ. Therapeutic potential of matrix metalloproteinase inhibitors in atherosclerosis. Exp Opin Invest Drugs. 2000;9:993–1007. doi: 10.1517/13543784.9.5.993. [DOI] [PubMed] [Google Scholar]
  • 8.Willenbrock F, Murphy G. Structure-function relationship in the tissue inhibitors of metalloproteinases. Am J Respir Crit Care Med. 1994;150:S165–70. doi: 10.1164/ajrccm/150.6_Pt_2.S165. [DOI] [PubMed] [Google Scholar]
  • 9.Baker AH, Zaltsman AB, George SJ, Newby AC. Divergent effects of tissue inhibitor of metalloproteinase-1,-2,-3 overexpression on rat vascular smooth muscle cell invasion, proliferation and death in vitro. TIMP-3 promotes apoptosis. J Clin Invest. 1998;101:1478–87. doi: 10.1172/JCI1584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.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;298:29–32. doi: 10.1016/0014-5793(92)80015-9. [DOI] [PubMed] [Google Scholar]
  • 11.Stetler-Stevenson WG, Bersch N, Golde DW. Tissue inhibitor of metalloproteinase-2 (TIMP-2) has erythroidpotentiating activity. FEBS Lett. 1992;296:231–4. doi: 10.1016/0014-5793(92)80386-u. [DOI] [PubMed] [Google Scholar]
  • 12.Duffy MJ, Maguire TM, Hill A, McDermott E, O'Higgins N. Metalloproteinases: role in breast carcinogenesis, invasion and metastasis. Brest Cancer Res. 2000;2:252–7. doi: 10.1186/bcr65. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Matrisian LM. Extracellular proteinases in malignancy. Current Biol. 1999;9:R776–8. doi: 10.1016/S0960-9822(00)80011-1. [DOI] [PubMed] [Google Scholar]
  • 14.McCawely LJ, Matrisian LM. Matrix metalloproteinases: multifunctional contributors to tumour progression. Mo Med Today. 2000;6:149–56. doi: 10.1016/s1357-4310(00)01686-5. [DOI] [PubMed] [Google Scholar]
  • 15.McCawely LJ, Matrisian LM. Matrix metalloproteinase: they're not just for matrix anymore! Curr Opin Cell Biol. 2001;13:534–40. doi: 10.1016/s0955-0674(00)00248-9. [DOI] [PubMed] [Google Scholar]
  • 16.Foda HD, Zucker S. Matrix metalloproteinase in cancer invasion, metastasis and angiogenesis. Drug Discovery Today. 2001;6:478–82. doi: 10.1016/s1359-6446(01)01752-4. [DOI] [PubMed] [Google Scholar]
  • 17.Fuberg AH, Ambrosone CB. Molecular epidemiology, biomarkers and cancer prevention. Trends in Molec Med. 2001;7:517–521. doi: 10.1016/s1471-4914(01)02162-1. [DOI] [PubMed] [Google Scholar]
  • 18.Giannelli G, Erriquez R, Fransvea E, Daniele A, Trerotoli P, Schittulli F, Grano M, Quaranta M, Antonaci S. Proteolytic imbalance is reversed after therapeutic surgery in breast cancer patients. Int J Cancer. 2004;109:782–5. doi: 10.1002/ijc.20009. [DOI] [PubMed] [Google Scholar]
  • 19.Würtz SØ, Schrohl A-S, Sørensen NM, Lademann U, Christensen IJ, Mouridsen H, Brünner N. Tissue inhibitor of metalloproteinases-1 in breast cancer. Endocrine-Related Cancer. 2005;12:125–227. doi: 10.1677/erc.1.00719. [DOI] [PubMed] [Google Scholar]
  • 20.Kuvaja P, Talvensaari-Mattila A, Paakko P, Turpeenniemi-Hujanen T. The absence of immunoreactivity for tissue inhibitor of metalloproteinase-1 (TIMP-1), but not for TIMP-2, protein is associated with the favorable prognossis in aggressive breast carcinoma. Oncology. 2005;68:196–203. doi: 10.1159/000086774. [DOI] [PubMed] [Google Scholar]
  • 21.Miller WR, Ellis IO, Sainsbury JRC, Dixon JM. ABC of breast diseases: prognostic factors. BMJ. 1994;309:1573–6. doi: 10.1136/bmj.309.6968.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Sainsbury JRC, Anderson TJ, Morgan DAL. ABC of breast diseases. BMJ. 2000;321:745–50. doi: 10.1136/bmj.321.7263.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Bussolati G, Gugliotta P. Nonspecific staining of mast cells by avidin-biotin-peroxidase complexes (ABC) J Histochem Cytochem. 1983;31:1419–21. doi: 10.1177/31.12.6195216. [DOI] [PubMed] [Google Scholar]
  • 24.Elston CW, Ellis IO, Pinder SE. Pathological prognostic factors in breast cancer. Crit Rev Oncol Hematol. 1999;31:209–23. doi: 10.1016/s1040-8428(99)00034-7. [DOI] [PubMed] [Google Scholar]
  • 25.Matache C, Stefanescu M, Dragomir C, Tanaseanu S, Onu A, Ofiteru A, Szegli G. Matrix Metalloproteinase-9 and its natural inhibitor TIMP-1 expressed or secreted by peripheral blood mononuclear cells from patients with systemic lupus erythematosus. Journal of Autoimmunity. 2003;20:323–31. doi: 10.1016/s0896-8411(03)00037-4. [DOI] [PubMed] [Google Scholar]
  • 26.Scorilas A, Karameris A, Arnogiannaki N, Ardavanis A, Bassilopoulos P, Trangas T, Talieri M. Overexpression of matrix-metalloproteinase-9 in human breast cancer: a potent favourable indicator in node-negative patients. Br J Cancer. 2001;84:1488–96. doi: 10.1054/bjoc.2001.1810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Liu SC, Yang SF, Yeh KT, Yeh CM, Chiou HL, Lee CY, Chou MC, Hsieh YS. Relationships between the level of matrix metalloproteinase-2 and tumor size of breast cancer. 2006;366:243–8. doi: 10.1016/j.cca.2006.02.026. [DOI] [PubMed] [Google Scholar]
  • 28.Remacle AG, Noel A, Duggan C, McDermott E, O'Higgins N, Foidart JM, Duffy MJ. Assay of matrix metalloproteinases types 1, 2, 3, and 9 in breast cancer. Brit J Cancer. 1998;77:926–31. doi: 10.1038/bjc.1998.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Nakopoulou L, Tsirmpa I, Alexandrou P, Louvrou A, Ampela C, Markaki S, Davaris PS. MMP-2 protein in invasive breast cancer and the impact of MMP-2/TIMP-2 phenotype on overall survival. Breast Cancer Res Treat. 2003;77:145–55. doi: 10.1023/a:1021371028777. [DOI] [PubMed] [Google Scholar]
  • 30.Hanemaaijer R, Verheijen JH, Maguire TM, Visser H, Toet K, McDermott E, O'Higgins N, Duffy MJ. Increased Gelatinase-A and Gelatinase-B activities in malignant vs benign breast tumors. Int J Cancer. 2000;86:204–7. doi: 10.1002/(sici)1097-0215(20000415)86:2<204::aid-ijc9>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  • 31.Wang HY, Zhang XB, Wang M. Expression of matrix metalloproteinase 9 (MMP-9) and laminin-receptor in breast carcinoma and their correlation with tumor metastasis and prognosis. Ai Zheng. 2003;22:529–32. [PubMed] [Google Scholar]
  • 32.Nakopoulou L, Katsarou S, Giannopoulou I, Alexandrou P, Tsirmpa I, Panayotopoulou E, Mavrommatis J, Keramopoulos A. Correlation of tissue inhibitor of Metalloproteinase-2 with proliferative activity and patients' survival in breast cancer. Mod Pathol. 2002;15:26–34. doi: 10.1038/modpathol.3880486. [DOI] [PubMed] [Google Scholar]
  • 33.Jones JL, Glynn P, Walker RA. Expression of MMP-2 and MMP-9, their inhibitors, and the activator MT1-MMP in primary breast carcinomas. J Pathol. 1999;189:161–8. doi: 10.1002/(SICI)1096-9896(199910)189:2<161::AID-PATH406>3.0.CO;2-2. [DOI] [PubMed] [Google Scholar]
  • 34.Garbett EA, Reed MWR, Stephenson TJ, Brown NJ. Proteolysis in human breast cancer. J Clin Pathol: Mol Pathol. 2000;53:99–106. doi: 10.1136/mp.53.2.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Iwata H, Kobayashi S, Iwase H, Masaoka A, Fujimoto N, Okada Y. Production of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human breast carcinomas. Jpn J Cancer Res. 1996;87:602–11. doi: 10.1111/j.1349-7006.1996.tb00266.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.McCarthy K, Maguire T, McGreal G, McDermott E, O'Higgins N, Duffy MJ. High levels of tissue inhibitor of metalloproteinase-1 predict poor outcome in patients with breast cancer. Int J Cancer. 1999;84:44–8. doi: 10.1002/(sici)1097-0215(19990219)84:1<44::aid-ijc9>3.0.co;2-p. [DOI] [PubMed] [Google Scholar]
  • 37.Fan SQ, Wei QY, Li MR, Zhang LQ, Liang QC. Expression and clinical significance of MMP-2, MMP-9, TIMP-1 and TIMP-2 in breast carcinoma. Ai Zheng. 2003;22:968–73. [PubMed] [Google Scholar]
  • 38.Rha SY, Yang WI, Kim JH, Roh JK, Min JS, Lee KS, Kim BS, Chung HC. Different expression patterns of MMP-2 and MMP-9 in breast cancer. Oncol Rep. 1998;5:875–9. doi: 10.3892/or.5.4.875. [DOI] [PubMed] [Google Scholar]
  • 39.Onisto M, Riccio MP, Scannapieco P, Caenazzo C, Griggio L, Spina M, Stetler-Stevenson WG, Garbisa S. Gelatinase A/TIMP-2 imbalance in lymph-node-positive breast carcinomas, as measured by RT-PCR. Int J Cancer. 1995;63:621–6. doi: 10.1002/ijc.2910630504. [DOI] [PubMed] [Google Scholar]
  • 40.Xu X, Wang Y, Chen Z, Sternlicht MD, Hidalgo M, Steffensen B. Matrix metalloproteinase-2 contributes to cancer cell migration on collagen. Cancer Res. 2005;65:130–6. [PubMed] [Google Scholar]
  • 41.Baker EA, Stephenson TJ, Reed MWR, Brown NJ. Expression of proteinases and inhibitors in human breast cancer progression and survival. J Clin Pathol: Mol Pathol. 2002;55:300–4. doi: 10.1136/mp.55.5.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Struse K, Audretsch W, Rezai M, Pott G, Bojar H. The estrogen receptor paradox in breast cancer: Association of high receptor concentrations with reduced overall survival. Breast J. 2000;6:115–25. doi: 10.1046/j.1524-4741.2000.99060.x. [DOI] [PubMed] [Google Scholar]
  • 43.Hou YF, Yuan ST, Li HC, Wu J, Lu JS, Liu G, Lu LJ, Shen ZZ, Ding J, Shao ZM. ERβ exerts multiple stimulative effects on human breast carcinoma cells. Oncogene. 2004;23:5799–806. doi: 10.1038/sj.onc.1207765. [DOI] [PubMed] [Google Scholar]
  • 44.Spencer F, Chi L, Zhu MX. Time-dependent relationship between the estrogen receptors and the matrix metalloproteinases following deciduoma induction in rats. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1998;120:283–8. doi: 10.1016/s0742-8413(98)10006-3. [DOI] [PubMed] [Google Scholar]
  • 45.Crowe DL, Brown TN. Transcriptional inhibition of matrix metalloproteinase 9 (MMP-9) activity by a c-fos/estrogen receptor fusion protein is mediated by the proximal AP-1 site of the MMP-9 promoter and correlates with reduced tumor cell invasion. Neoplasia. 1999;1:368–72. doi: 10.1038/sj.neo.7900041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Potier M, Elliot SJ, Tack I, Lenz O, Striker GE, Striker LJ, Karl M. Expression and regulation of estrogen receptors in mesangial cells: influence on matrix metalloproteinase-9. J Am Soc Nephrol. 2001;12:241–51. doi: 10.1681/ASN.V122241. [DOI] [PubMed] [Google Scholar]
  • 47.Razandi M, Pedram A, Park ST, Levin ER. Proximal events in signalling by plasma membrane estrogen receptors. J Biol Chem. 2003;24:2710–2. doi: 10.1074/jbc.M205692200. [DOI] [PubMed] [Google Scholar]
  • 48.Nakopoulou L, Michalopoulou A, Giannopoulou I, Tzonou A, Keramopoulos A, Lazaris AC, Davaris PS. Bcl-2 protein expression is associated with a prognostically favourable phenotype in breast cancer irrespective of p53 immunostaining. Histopathology. 1999;34:310–9. doi: 10.1046/j.1365-2559.1999.00627.x. [DOI] [PubMed] [Google Scholar]
  • 49.Oliver L, Tremblais K, Guriec N, Martin S, Meflah K, Menanteau J, Vallette FM. Influence of bcl-2-related proteins on matrix metalloproteinase expression in a rat glioma cell line. Biochem Biophys Res Commun. 2000;273:411–6. doi: 10.1006/bbrc.2000.2952. [DOI] [PubMed] [Google Scholar]
  • 50.Ricca A, Biroccio A, Del Bufalo D, Mackay AR, Santoni A, Cippitelli M. bcl-2 over-expression enhances NF-k B activity and induces mmp-9 transcription in human MCF7 (ADR) breast-cancer cells. Int J Cancer. 2000;86:188–96. doi: 10.1002/(sici)1097-0215(20000415)86:2<188::aid-ijc7>3.0.co;2-w. [DOI] [PubMed] [Google Scholar]
  • 51.Liu XW, Taube ME, Jung KK, Dong Z, Lee YJ, Roshy S, Sloane BF, Fridman R, Kim HR. Tissue inhibitor of metalloproteinase-1 protects human breast epithelial cells from extrinsic cell death: a potential oncogenic activity of tissue inhibitor of metalloproteinase-1. Cancer Res. 2005;65:898–906. [PubMed] [Google Scholar]
  • 52.Martinez-Arribas F, Nunez-Villar MJ, Lucas AR, Sanchez J, Tejerina A, Schneider J. Immunofluorometric study of Bcl-2 and Bax expression in clinical fresh tumor samples from breast cancer patients. Anti Cancer Res. 2003;23:565–8. [PubMed] [Google Scholar]
  • 53.Fitzgibbons PL, Page DL, Weaver D, Thor AD, Allred DC, Clark GM, Ruby SG, O'Malley F, Simpson JF, Connolly JL, Hayes DF, Edge SB, Lichter A, Schnitt SJ. Prognostic factors in breast cancer. College of American Pathologists Consensus Statement 1999. Arch Pathol Lab Med. 2000;124:966–78. doi: 10.5858/2000-124-0966-PFIBC. [DOI] [PubMed] [Google Scholar]

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