Characteristics of a 95-kDa matrix metalloproteinase produced by mammary carcinoma cells

Lyons, J. Guy; Birkedal-Hansen, B; Moore, William G. I.; O'Grady, Robert L.; Birkedal‐Hansen, Henning

doi:10.1021/bi00220a001

Cited by 86 publications

(70 citation statements)

References 35 publications

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“…Fibrinogen is degraded by MMP-9 (20)(21)(22). As a result, integrinmediated interaction may switch from support of firm adhesion to dynamic migration on the modified substrate (17).…”

Section: Discussionmentioning

confidence: 99%

“…MMP-9 may affect fibrinogen-directed breast cancer cell migration by modifying the substrate and converting it into a promigratory format (20)(21)(22). The 82-kDa metalloproteinase produced by breast cancer cells expressing activated ␣v␤3 degraded fibrinogen, as shown by zymography (Fig.…”

Section: Breast Cancer Cells With Activated ␣V␤3 Produce the Mature Fmentioning

confidence: 96%

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Activated integrin αvβ3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells

Rolli

Fransvea

Pilch

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

287

250

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Expression of adhesion receptor integrin ␣v␤3 in an activated functional form strongly promotes metastasis in human breast cancer cells. Here, we report that ␣v␤3 cooperates with matrix metalloproteinase type 9 (MMP-9) in breast cancer cell migration. This cooperation is regulated by the activation state of the integrin. Expression of activated ␣v␤3 in metastatic variants of MDA-MB 435 human breast cancer cells and primary metastatic cells from breast cancer patients strongly enhanced migration toward vitronectin and fibrinogen. This enhancement was mediated by a soluble factor produced by breast cancer cells expressing activated ␣v␤3. When transferred, this factor also up-regulated ␣v␤3-dependent migration of breast cancer cells that express the nonactivated integrin. The factor was identified as metalloproteinase MMP-9. Whereas all tested breast cancer cell variants produced latent MMP-9, only those with activated ␣v␤3 produced the mature form of this metalloproteinase. Recombinant mature MMP-9, but not latent MMP-9 or either form of MMP-2, enhanced ␣v␤3-dependent breast cancer cell migration. The migratory response was inhibited by tissue inhibitors of metalloproteinase or when MMP-9 was depleted from the inducing supernatants. The results indicate a causal relationship between the expression of activated integrin ␣v␤3 and production of enzymatically active MMP-9 in metastatic breast cancer cells. These molecules cooperate to enhance breast cancer cell migration toward specific matrix proteins, and this may contribute to the strongly enhanced metastatic capacity of breast cancer cells that express activated ␣v␤3.M etastasis is the primary cause of death in breast cancer patients. Metastatic dissemination depends on tumor cell adhesion, migration, and invasion. These steps involve integrins, a family of transmembrane adhesion receptors, composed of noncovalently linked ␣ and ␤ subunits (1). Integrins are known to exist in distinct states of activation, and these determine integrin functionality and affinity for ligands (2). For example, integrin activation controls which ligands are recognized, whether an integrin can support cell arrest under dynamic flow conditions or only stationary adhesion, and whether cells can migrate on or toward specific substrates (3). The importance of integrin activation has long been appreciated in leukocytes and platelets, where it controls inflammatory responses and thrombus formation (4, 5). Recent findings indicate that other cell types, such as endothelial cells and tumor cells, can also regulate their interaction with extracellular matrix proteins by integrin activation (6-9). This regulation may help to control angiogenesis and tumor metastasis.In breast and ovarian cancer, as well as in melanoma and glioma, malignant progression is associated with expression of tumor cell integrin ␣v␤3 (10-14). We recently found that ␣v␤3 can exist in human breast cancer cells in an activated or a nonactivated functional state. Only the activated state supports breast cancer cell arrest du...

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“…Fibrinogen is degraded by MMP-9 (20)(21)(22). As a result, integrinmediated interaction may switch from support of firm adhesion to dynamic migration on the modified substrate (17).…”

Section: Discussionmentioning

confidence: 99%

Section: Breast Cancer Cells With Activated ␣V␤3 Produce the Mature Fmentioning

confidence: 96%

Activated integrin αvβ3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells

Rolli

Fransvea

Pilch

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

287

250

View full text Add to dashboard Cite

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“…Cleavage of this bond triggers a change in proMMP-9 that renders the Arg-87-Phe-88 amide bond susceptible to the second cleavage, resulting in conversion to an 82-kDa species (Ogata et al 1992: Fridman et al, 1995. ProMMP-9 can be activated autocatalytically by organomercurial compounds or trypsin in vitro (Wilhelm et al, 1989;Lyons et al, 1991). In the current study.…”

Section: Discussionmentioning

confidence: 99%

Unique activation of matrix metalloproteinase-9 within human liver metastasis from colorectal cancer

Zeng

Guillem²

1998

Br J Cancer

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Summary Expermental in vitro and animal data support an important role for matrix metalloproteinases (MMPs) in cancer invasion and metastasis via proteolytic degradation of the extracellular matrix (ECM). Our previous data have shown that MMP-9 mRNA is localized to the interface between liver metastasis and normal liver tissue, indicating that MMP-9 may play an important role in liver metastasis formation. In the present study, we analysed the cellular enzymatic expression of MMP-9 in 18 human colorectal cancer (CRC) liver metastasis specimens by enzyme-linked immunosorbent assay (ELISA) and zymography. ELISA anatysis reveals that the latent form of MMP-9 is present in both liver metastasis and paired adjacent normal liver tissue. The mean level of the latent form of MMP-9 is 580±270 ng per mg total tissue protein (mean ± s.e.) in liver metastasis vs 220 ± 90 in normal liver tissue. However, this difference is not significantly different (P = 0.26). Using gelatin zymography, the 92-kDa band representative of the latent form is present in both liver metastasis and normal liver tissue. However, the 82 kDa band, representive of the active form of MMP-9, was seen only in liver metastasis. This was confirmed by Westem blot analysis. Our observation of the unique presence of the active form of MMP-9 within liver metastasis suggests that proMMP-9 activation may be a pivotal event during CRC liver metastasis formation. . emphasizing the clinical importance of elevated MMP-9 expression in primarx CRC.We hax-e previously demonstrated that MMP-9 mRRNA is localized in the interface between lixer metastasis and normal lixer tissue. indicating that MMP-9 may play an important role in lix er metastasis formation. To inxestigate further the significance of elexated MMP-9 RNA in liver metastasis formation. the present study examined MMP-9 enzy matic expression in CRC liver metastasis. MATERIALS AND METHODS

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“…ProMMP-9 has been purified from human (Hibbs et al, 1985) and pig neutrophils (Murphy et al, 1989a), and from the culture medium of several transformed human cells (Wilhelm et al, 1989;Davis & Martin, 1990;Moll et al, 1990) and rat mammary carcinoma cells (Lyons et al, 1991). isolated from the transformed cells was shown to be noncovalently complexed with TIMP.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of matrix metalloproteinase 9 from U937 monocytic leukaemia and HT1080 fibrosarcoma cells

Morodomi

Osuga

Sasaguri

et al. 1992

Biochemical Journal

185

126

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603The precursor of matrix metalloproteinase 9 (proMMP-9), also known as '92 kDa progelatinase/type IV procollagenase', was purified from the conditioned medium of U937 monocytic leukaemia and HT1080 fibrosarcoma cell lines stimulated with phorbol 12-myristate 13-acetate. ProMMP-9 in these culture media is non-covalently complexed with the 29 kDa tissue inhibitor of metalloproteinases (TIMP), but free proMMP-9 was separated from the TIMP-proMMP-9 complex by chromatography on Green A Dyematrex gel. The final product was homogeneous on SDS/PAGE, with a molecular mass of 88 kDa without reduction and 92 kDa with reduction. Treatment of proMMP-9 with 4-aminophenylmercuric acetate converted the 88 kDa precursor into 80 kDa and 68 kDa forms. Gelatin-containing zymographic analysis showed zones of lysis associated with all three species. However, only the 68 kDa species was shown to be catalytically active by its ability to bind to a2-macroglobulin. In the presence of an equimolar amount of TIMP, only the 80 kDa species was generated by treatment with 4-aminophenylmercuric acetate, but no enzyme activity was detected. This indicates that TIMP binds to the 80 kDa intermediate and inhibits the generation of the active 68 kDa species. Eight endopeptidases (trypsin, chymotrypsin, plasmin, plasma kallikrein, thrombin, cathepsin G, neutrophil elastase and thermolysin) were tested for their ability to activate proMMP-9. Of them, trypsin was the most effective activator of proMMP-9. Only partial activation (10-30%) was observed with plasmin, cathepsin G and chymotrypsin. The active forms generated by trypsin were identified as 80 kDa, 74 kDa and 66 kDa by their abilities to bind to a2-macroglobulin. In the presence of an equimolar amount of TIMP, proMMP-9 was also converted into the same molecular-mass species by trypsin, but they were not proteolytically active. This suggests activated MMP-9 is inhibited by TIMP. Activated MMP-9 digested gelatin, type-V collagen, reduced carboxymethylated transferrin and, to a lesser extent, type-IV collagen and laminin A chain. The specific activity against gelatin was estimated to be 15000 units/mg (1 unit = 1 ,ug of gelatin degraded/min at 37°C) by titration with a2-macroglobulin. Comparative studies on digestion of gelatin and collagen types IV and V by MMP-9 and MMP-2 indicated that both enzymes degrade these substrates into similar fragments. However, the susceptibilities of laminin, fibronectin and reduced carboxymethylated transferrin to these two MMPs were sufficiently different to indicate differences in substrate specificities between these two closely related proteinases.

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Characteristics of a 95-kDa matrix metalloproteinase produced by mammary carcinoma cells

Cited by 86 publications

References 35 publications

Activated integrin αvβ3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells

Activated integrin αvβ3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells

Unique activation of matrix metalloproteinase-9 within human liver metastasis from colorectal cancer

Purification and characterization of matrix metalloproteinase 9 from U937 monocytic leukaemia and HT1080 fibrosarcoma cells

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