Posttranscriptional Stimulation of Endothelial Cell Matrix Metalloproteinases 2 and 1 by Endothelioma Cells

Taraboletti, Giulia; Sonzogni, Laura; Vergani, Veronica; Hosseini, Ghamartaj; Ceruti, Roberta; Ghilardi, Carmen; Bastone, Antonio; Toschi, Elena; Borsotti, Patrizia; Scanziani, Eugenio; Giavazzi, Raffaella; Pepper, Michael Sean; Stetler-Stevenson, William G.; Bani, Maria Rosa

doi:10.1006/excr.2000.4936

Cited by 42 publications

(28 citation statements)

References 34 publications

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“…This is borne out by the finding that both inactive and activated MMP-2 and MMP-9 accumulate in the cytosol of human endothelial cells, 16 confirming the existence of intracellular storage compartments and activation mechanisms. In addition, our previous finding that angiogenesis-stimulating factors rapidly induced secretion of MMP-2 by endothelial cells, independently of protein synthesis, 17 further suggests that endothelial cell proteolytic activity can be stimulated by quick mobilization of intracellular MMP pools.…”

Section: Discussionmentioning

confidence: 81%

“…16 We have previously reported that MMP production by endothelial cells was increased without de novo synthesis and was prevented by inhibitors of secretion, indicating that protease production may indeed be regulated at the level of secretion in these cells. 17 This study was designed to investigate whether shedding MMPs as membrane vesicle components might be a mechanism of rapid secretion of proteases during angiogenesis. We investigated whether endothelial cells shed MMP-containing vesicles, whether this release was modulated by angiogenic factors, and whether MMP-containing vesicles played a role in endothelial cell functions related to angiogenesis.…”

mentioning

confidence: 99%

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Shedding of the Matrix Metalloproteinases MMP-2, MMP-9, and MT1-MMP as Membrane Vesicle-Associated Components by Endothelial Cells

Taraboletti

D’Ascenzo

Borsotti

et al. 2002

The American Journal of Pathology

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509

381

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Production of matrix-degrading proteases, particularly matrix metalloproteinases (MMPs), by endothelial cells is a critical event during angiogenesis, the process of vessel neoformation that occurs in normal and pathological conditions. MMPs are known to be highly regulated at the level of synthesis and activation, however, little is known about the regulation of MMP secretion by endothelial cells. We found that cultured human umbilical vein endothelial cells shed vesicles (300 to 600 nm) originating from localized areas of the cell plasma membrane, as revealed by ultrastructural analysis. Normal and reverse zymography, Western blot, and immunogold analyses of the vesicles showed two gelatinases, MMP-2 and MMP-9, in both the active and proenzyme forms, the MT1-MMP proenzyme located on the external side of the vesicle membrane and the two inhibitors TIMP-1 and TIMP-2. Serum and the angiogenic factors, fibroblast growth factor-2 and vascular endothelial growth factor, stimulated the shedding of MMPs as vesicle components. Shedding the vesicle was rapid, as it was already completed after 4 hours. Addition of shed vesicles to human umbilical vein endothelial cells resulted in autocrine stimulation of invasion through a layer of reconstituted basement membrane (Matrigel) and cord formation on Matrigel. We conclude that endothelial cells shed MMP-containing vesicles and this may be a mechanism for regulating focalized proteolytic activity vital to invasive and morphogenic events during angiogenesis. (Am J Pathol 2002, 160:673-680)

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Section: Discussionmentioning

confidence: 81%

mentioning

confidence: 99%

Shedding of the Matrix Metalloproteinases MMP-2, MMP-9, and MT1-MMP as Membrane Vesicle-Associated Components by Endothelial Cells

Taraboletti

D’Ascenzo

Borsotti

et al. 2002

The American Journal of Pathology

Self Cite

509

381

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“…34 Confluent monolayers of hTERT-HDLEC or HMEC-1 were washed with PBS and the cells incubated in their corresponding serum-and cytokine-free media. After a 15 hour incubation at 37°C, conditioned media were collected, supplemented with 0.5 mmol/L phenylmethylsulfonyl fluoride (PMSF) and 15 mmol/L N-(2-hydroxyethyl)piperazine-NЈ-(2 ethanesulfonic acid) (HEPES), centrifuged at 340 ϫ g for 5 minutes, and the resulting supernatants were stored at Ϫ80°C until use.…”

Section: Zymography and Reverse Zymographymentioning

confidence: 99%

Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span

Nisato

Buser

et al. 2004

The American Journal of Pathology

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The study of lymphatic endothelial cells and lymphangiogenesis has, in the past, been hampered by the lack of lymphatic endothelial-specific markers. The recent discovery of several such markers has permitted the isolation of lymphatic endothelial cells (LECs) from human skin. However, cell numbers are limited and purity is variable with the different isolation procedures. To overcome these problems, we have transfected human dermal microvascular endothelial cells (HDMVECs) with a retrovirus containing the coding region of human telomerase reverse transcriptase (hTERT), and have produced a cell line, hTERT-HDLEC, with an extended lifespan. hTERT-HDLEC exhibit a typical cobblestone morphology when grown in culture, are contact-inhibited, and express endothelial cell-specific markers. hTERT-HDLEC also express the recognized lymphatic markers, Prox-1, LYVE-1 and podoplanin, as well as integrin ␣9, but do not express CD34. They also form tube-like structures in three-dimensional collagen gels when stimulated with vascular endothelial growth factors -A and -C. Based on these currently recognized criteria, these cells are LEC. Surprisingly, we also found that the widely studied HMEC-1 cell line expresses recognized lymphatic markers; however, these cells are also CD34-positive. In summary, the ectopic expression of hTERT increases the life span of LECs and does not affect their capacity to form tubelike structures in a collagen matrix. The production and characterization of hTERT-HDLEC will facilitate the study of the properties of lymphatic endothelium in vitro. The blood and lymphatic vascular systems are anatomically and histologically closely related but distinct, and do not anastomose except at the level of the main lymphatic ducts. The blood vascular system supplies nutrients and oxygen to the body while the lymphatic vasculature is crucial for immune cell traffic and the uptake of extracellular fluid. The formation of new blood vessels from pre-existing blood vessels (angiogenesis) occurs in physiological (eg, postnatal growth, inflammation, and tissue repair) and pathological (eg, tumor recruitment of blood vessels) settings. Lymphangiogenesis is the formation of new lymphatic vessels from pre-existing lymphatic capillaries, and occurs in similar settings. Blood endothelial cells (BECs) and lymphatic endothelial cells (LECs) are quiescent cells that line the lumina of blood and lymphatic vessels.The lymphatic system has traditionally been overshadowed by the greater emphasis placed on the blood vascular system. This has been due in part to the absence of suitable markers that distinguish lymphatic from blood vascular endothelium. In the past few years, this limitation has been overcome. Lymphatic markers include LYVE-1, a lymphatic endothelial receptor for the extracellular matrix/lymphatic fluid glycosaminoglycan hyaluronan; 1 Prox-1, a homeobox gene product involved in regulating early lymphatic development; 2 podoplanin, a glomerular podocyte membrane mucoprotein; 3 and vascular endothelial growth factor re...

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“…For immunohistochemical analysis of CD31 (a speci®c marker of endothelial cells), sections were immunostained with a rat anti-mouse CD31 monoclonal antibody (Vecchi et al, 1994), according to the described procedure (Taraboletti et al, 2000). Microvessel count was carried out on three ®elds (6250) chosen within the highest vascularized areas.…”

Section: In Vivo Staining Of Tumor Vessels and Apoptotic Cellsmentioning

confidence: 99%

“…For the latter, the procedure described in (Taraboletti et al, 2000) was used, except that the antibody was diluted 1 : 6000 and that sections were pre-incubated with goat serum. In both assays, count of apoptotic cells was carried out on 10 random ®elds (6400).…”

Section: In Vivo Staining Of Tumor Vessels and Apoptotic Cellsmentioning

confidence: 99%

p73 overexpression increases VEGF and reduces thrombospondin-1 production: implications for tumor angiogenesis

et al. 2001

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Posttranscriptional Stimulation of Endothelial Cell Matrix Metalloproteinases 2 and 1 by Endothelioma Cells

Cited by 42 publications

References 34 publications

Shedding of the Matrix Metalloproteinases MMP-2, MMP-9, and MT1-MMP as Membrane Vesicle-Associated Components by Endothelial Cells

Shedding of the Matrix Metalloproteinases MMP-2, MMP-9, and MT1-MMP as Membrane Vesicle-Associated Components by Endothelial Cells

Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span

p73 overexpression increases VEGF and reduces thrombospondin-1 production: implications for tumor angiogenesis

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