Contact of high-invasive, but not low-invasive, melanoma cells to native collagen I induces the release of mature cathepsin B.
Metastasis of malignant tumor cells involves cell-cell and cell-matrix interactions, which regulate the expression and localization of proteolytic enzymes. In the present study, we investigated the expression and localization of the lysosomal cysteine proteinase cathepsin B and its natural inhibitors cystatin A, B and C in high- (MV3), intermediate- (SKmel28) and low-invasive (SKmel23, WM164) human melanoma cell lines grown on plastic or in contact with monomeric or fibrillar collagen type I. Neither the transcript levels of cathepsin B nor those of the natural inhibitors, cystatin B and C, were altered by the interaction of melanoma cells with collagen type I. However, protein expression and cellular localization of cathepsin B and its inhibitors were markedly affected. In contrast to low-invasive cells, high-invasive cells constitutively released procathepsin B when cultured on plastic. In addition, contact of invasive cells with fibrillar collagen type I resulted in the release of both mature forms of the protease. Perturbation studies using inhibitory antibodies against the beta1 subunit of the integrin receptor indicated a role for the beta1 integrin receptor family in the regulation of cathepsin B release. Cystatin B protein expression was much lower in high-invasive cells in both culture conditions, when compared to low-invasive cells. Cystatin C expression was comparable in all cells, but cell contact to fibrillar collagen type I induced its expression. These results strongly implicate a pivotal role of cell-matrix interactions for the regulation of cathepsin B localization and activity in melanoma cells.
Klose
Anke
A
Department of Dermatology, Center for Molecular Medicine (CMMC), University of Cologne, Cologne, Germany.
Wilbrand-Hennes
Astrid
A
Zigrino
Paola
P
Weber
Ekkehard
E
Krieg
Thomas
T
Mauch
Cornelia
C
Hunzelmann
Nicolas
N
D002401:Cathepsin B / Q000032:analysis / Q000096:biosynthesis*; D021982:Cell-Matrix Junctions / Q000502:physiology*; D024042:Collagen Type I / Q000502:physiology*; D055313:Cystatin B; D055316:Cystatin C; D015891:Cystatins / Q000096:biosynthesis / Q000502:physiology; D020869:Gene Expression Profiling; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000473:pathology*; D012878:Skin Neoplasms / Q000235:genetics / Q000473:pathology*; D014407:Tumor Cells, Cultured
C525030:CST3 protein, human; C524994:CSTB protein, human; D024042:Collagen Type I; D055316:Cystatin C; D015891:Cystatins; D055313:Cystatin B; D002401:Cathepsin B