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TNF-alpha increases human melanoma cell invasion and migration in vitro: the role of proteolytic enzymes.

Katerinaki E, Evans GS, Lorigan PC, MacNeil S - Br. J. Cancer (2003)

Bottom Line: In this study, we extend these investigations asking specifically whether the TNF-alpha effect on cell invasion and migration involves activation of proteolytic enzymes.We examined the effect of TNF-alpha on melanoma expression/activation of type IV gelatinases matrix metalloproteinases 2 and 9 (MMPs -2 and -9) and general proteolytic enzymes.These findings suggest that the promigratory and proinvasive effect of TNF-alpha on this melanoma cell line may be mediated to some extent by induction of localised cell membrane-bound degradative enzyme activity, which is not readily detected in biochemical assays.

View Article: PubMed Central - PubMed

Affiliation: Section of Medicine, Division of Clinical Sciences, Northern General Hospital, Herries Road, Sheffield S5 7AU, UK.

ABSTRACT
Inflammatory mediators have been reported to promote malignant cell growth, invasion and metastatic potential. More specifically, we have recently reported that tumour necrosis factor alpha (TNF-alpha) increases melanoma cell attachment to extracellular matrix (ECM) substrates and invasion through fibronectin. In this study, we extend these investigations asking specifically whether the TNF-alpha effect on cell invasion and migration involves activation of proteolytic enzymes. We examined the effect of TNF-alpha on melanoma expression/activation of type IV gelatinases matrix metalloproteinases 2 and 9 (MMPs -2 and -9) and general proteolytic enzymes. Stimulation with TNF-alpha significantly increased both melanoma cell migration at 24 h (+21%) and invasion through fibronectin (+35%) but did not upregulate/activate the expression of latent MMP-2 constitutively produced by these cells and did not upregulate their general protease activity. However, the increased cell migration and invasion through fibronectin observed following stimulation with TNF-alpha were inhibited by the general protease inhibitor alpha(2) macroglobulin. These findings suggest that the promigratory and proinvasive effect of TNF-alpha on this melanoma cell line may be mediated to some extent by induction of localised cell membrane-bound degradative enzyme activity, which is not readily detected in biochemical assays.

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Representative gelatin zymograph of control and TNF-α-stimulated HBL melanoma cell extracts and cell conditioned medium. Cells constitutively expressed pro-MMP-2 (66.3 kDa band) in the culture medium and no upregulation or activation of the latent enzyme was observed after preincubation with TNF-α at 200 U ml−1. Lanes 1 and 2 represent internal human purified MMP-2 and MMP-9 standards.
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fig2: Representative gelatin zymograph of control and TNF-α-stimulated HBL melanoma cell extracts and cell conditioned medium. Cells constitutively expressed pro-MMP-2 (66.3 kDa band) in the culture medium and no upregulation or activation of the latent enzyme was observed after preincubation with TNF-α at 200 U ml−1. Lanes 1 and 2 represent internal human purified MMP-2 and MMP-9 standards.

Mentions: HBL cells were incubated with TNF-α 200 U ml−1for 24 h prior to cell extraction and collection of cell conditioned medium for gelatin zymography (results are shown in Figure 2Figure 2


TNF-alpha increases human melanoma cell invasion and migration in vitro: the role of proteolytic enzymes.

Katerinaki E, Evans GS, Lorigan PC, MacNeil S - Br. J. Cancer (2003)

Representative gelatin zymograph of control and TNF-α-stimulated HBL melanoma cell extracts and cell conditioned medium. Cells constitutively expressed pro-MMP-2 (66.3 kDa band) in the culture medium and no upregulation or activation of the latent enzyme was observed after preincubation with TNF-α at 200 U ml−1. Lanes 1 and 2 represent internal human purified MMP-2 and MMP-9 standards.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376936&req=5

fig2: Representative gelatin zymograph of control and TNF-α-stimulated HBL melanoma cell extracts and cell conditioned medium. Cells constitutively expressed pro-MMP-2 (66.3 kDa band) in the culture medium and no upregulation or activation of the latent enzyme was observed after preincubation with TNF-α at 200 U ml−1. Lanes 1 and 2 represent internal human purified MMP-2 and MMP-9 standards.
Mentions: HBL cells were incubated with TNF-α 200 U ml−1for 24 h prior to cell extraction and collection of cell conditioned medium for gelatin zymography (results are shown in Figure 2Figure 2

Bottom Line: In this study, we extend these investigations asking specifically whether the TNF-alpha effect on cell invasion and migration involves activation of proteolytic enzymes.We examined the effect of TNF-alpha on melanoma expression/activation of type IV gelatinases matrix metalloproteinases 2 and 9 (MMPs -2 and -9) and general proteolytic enzymes.These findings suggest that the promigratory and proinvasive effect of TNF-alpha on this melanoma cell line may be mediated to some extent by induction of localised cell membrane-bound degradative enzyme activity, which is not readily detected in biochemical assays.

View Article: PubMed Central - PubMed

Affiliation: Section of Medicine, Division of Clinical Sciences, Northern General Hospital, Herries Road, Sheffield S5 7AU, UK.

ABSTRACT
Inflammatory mediators have been reported to promote malignant cell growth, invasion and metastatic potential. More specifically, we have recently reported that tumour necrosis factor alpha (TNF-alpha) increases melanoma cell attachment to extracellular matrix (ECM) substrates and invasion through fibronectin. In this study, we extend these investigations asking specifically whether the TNF-alpha effect on cell invasion and migration involves activation of proteolytic enzymes. We examined the effect of TNF-alpha on melanoma expression/activation of type IV gelatinases matrix metalloproteinases 2 and 9 (MMPs -2 and -9) and general proteolytic enzymes. Stimulation with TNF-alpha significantly increased both melanoma cell migration at 24 h (+21%) and invasion through fibronectin (+35%) but did not upregulate/activate the expression of latent MMP-2 constitutively produced by these cells and did not upregulate their general protease activity. However, the increased cell migration and invasion through fibronectin observed following stimulation with TNF-alpha were inhibited by the general protease inhibitor alpha(2) macroglobulin. These findings suggest that the promigratory and proinvasive effect of TNF-alpha on this melanoma cell line may be mediated to some extent by induction of localised cell membrane-bound degradative enzyme activity, which is not readily detected in biochemical assays.

Show MeSH
Related in: MedlinePlus