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Titolo:
Imaging proteolysis by living human glioma cells
Autore:
Sameni, M; Dosescu, J; Sloane, BF;
Indirizzi:
Wayne State Univ, Dept Pharmacol, Sch Med, Detroit, MI 48201 USA Wayne State Univ Detroit MI USA 48201 col, Sch Med, Detroit, MI 48201 USA Wayne State Univ, Barbara Ann Karmanos Canc Inst, Sch Med, Detroit, MI 48201 USA Wayne State Univ Detroit MI USA 48201 nst, Sch Med, Detroit, MI 48201 USA
Titolo Testata:
BIOLOGICAL CHEMISTRY
fascicolo: 5, volume: 382, anno: 2001,
pagine: 785 - 788
SICI:
1431-6730(200105)382:5<785:IPBLHG>2.0.ZU;2-2
Fonte:
ISI
Lingua:
ENG
Soggetto:
CATHEPSIN-B EXPRESSION; BREAST-CANCER CELLS; EXTRACELLULAR-MATRIX; INVASION; PROGRESSION; ASSOCIATION;
Keywords:
cathepsin B; endocytosis; extracellular matrix; laser scanning confocal microscopy; lysosomal proteases; quenched fluorescent substrates;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
18
Recensione:
Indirizzi per estratti:
Indirizzo: Sloane, BF Wayne State Univ, Dept Pharmacol, Sch Med, Detroit, MI 48201 USA Wayne State Univ Detroit MI USA 48201 d, Detroit, MI 48201 USA
Citazione:
M. Sameni et al., "Imaging proteolysis by living human glioma cells", BIOL CHEM, 382(5), 2001, pp. 785-788

Abstract

Degradation of basement membrane is an essential step for tumor invasion. In order to study degradation in real time as well as localize the site of proteolysis, we have established an assay with living human cancer cells inwhich we image cleavage of quenched-fluorescent basement membrane type IV collagen (DQ-collagen IV). Accumulation of fluorescent products is imaged with a confocal microscope and localized by optically sectioning both the cells and the matrix on which they are growing. For the studies described here, we seeded U87 human glioma cells as either monolayers or spheroids on a 3-dimensional gelatin matrix in which DQ-collagen IV had been embedded. As early as 24 hours after plating as monolayers, U87 cells were present throughout the 3-dimensional matrix. Cells at all levels had accumulated fluorescent degradation products of DQ-collagen IV intracellularly within vesicles. Similar observations were made for U87 spheroids and the individual cellsmigrating from the spheroids into the gelatin matrix. Both the migrating cells and those within the spheroid contained fluorescent degradation products of DQ-collagen IV intracellularly within vesicles. Thus, glioma cells like breast cancer cells are able to degrade type IV collagen intracellularly, suggesting that this is an important pathway for matrix degradation.

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Documento generato il 11/07/20 alle ore 16:02:31