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Matrix metalloproteinase-1 contribution to sarcoma cell invasion.

Garamszegi N, Garamszegi SP, Scully SP - J. Cell. Mol. Med. (2012)

Bottom Line: Inhibition of prenylation by farnesyl transferase inhibitor (FTI-276) decreased extracellular MMP-1 and subsequently reduced invasiveness by 30%.Parallel, stable cell line RNAi knockdown of MMP-1 confirmed its role in cellular invasiveness.These results indicate that MMP-1 directional delivery to podia structures is involved in the invasive activity of sarcoma cells, and this process is prenylation sensitive.

View Article: PubMed Central - PubMed

Affiliation: Sarcoma Biology Laboratory of Sylvester Comprehensive Cancer Center, University of Miami, Miller School of Medicine, FL, USA. ngaramszegi@med.miami.edu

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Related in: MedlinePlus

MMP-1 directional trafficking into invadopodia regions. (A) 143B cells were seeded in 12-well plates at 3.0 χ 104 concentrations. The live cell-related matrix degradation and MMP1-DsRed fusion construct production was imaged between 24 and 36 hrs from the DNA transfection. Except the DsRed fusion protein (constant fluorescence signal), the DQ-collagen becomes fluorescent upon enzymatic cleavage and degradation (three representative fields are shown, bar: 20 μm). (B) Images of MMP-1-DsRed transfected live cells. The data show that all fluorescent signal (green, red, blue,) demonstrate partial co-localization (inset), with white pixels where all three is simultaneously present (bars 20 μm).
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fig04: MMP-1 directional trafficking into invadopodia regions. (A) 143B cells were seeded in 12-well plates at 3.0 χ 104 concentrations. The live cell-related matrix degradation and MMP1-DsRed fusion construct production was imaged between 24 and 36 hrs from the DNA transfection. Except the DsRed fusion protein (constant fluorescence signal), the DQ-collagen becomes fluorescent upon enzymatic cleavage and degradation (three representative fields are shown, bar: 20 μm). (B) Images of MMP-1-DsRed transfected live cells. The data show that all fluorescent signal (green, red, blue,) demonstrate partial co-localization (inset), with white pixels where all three is simultaneously present (bars 20 μm).

Mentions: General paradigm is that MMP-1 secreted and deposited in the extracellular matrix as an inactive zymogen by various cell types where it is subsequently activated [47]. To investigate further if MMP-1 cellular production and localization can overlap with the underlying collagen matrix degradation beneath the cells, live cell fluorescent imaging in combination with DQ-collagen, MMP1-DsRed fusion protein expression and MMP-1 cleavage specific Fluorogenic Substrate-III (Fig. 4A and B; Refs. [48, 49]) were used in invadopodia assays.


Matrix metalloproteinase-1 contribution to sarcoma cell invasion.

Garamszegi N, Garamszegi SP, Scully SP - J. Cell. Mol. Med. (2012)

MMP-1 directional trafficking into invadopodia regions. (A) 143B cells were seeded in 12-well plates at 3.0 χ 104 concentrations. The live cell-related matrix degradation and MMP1-DsRed fusion construct production was imaged between 24 and 36 hrs from the DNA transfection. Except the DsRed fusion protein (constant fluorescence signal), the DQ-collagen becomes fluorescent upon enzymatic cleavage and degradation (three representative fields are shown, bar: 20 μm). (B) Images of MMP-1-DsRed transfected live cells. The data show that all fluorescent signal (green, red, blue,) demonstrate partial co-localization (inset), with white pixels where all three is simultaneously present (bars 20 μm).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3823085&req=5

fig04: MMP-1 directional trafficking into invadopodia regions. (A) 143B cells were seeded in 12-well plates at 3.0 χ 104 concentrations. The live cell-related matrix degradation and MMP1-DsRed fusion construct production was imaged between 24 and 36 hrs from the DNA transfection. Except the DsRed fusion protein (constant fluorescence signal), the DQ-collagen becomes fluorescent upon enzymatic cleavage and degradation (three representative fields are shown, bar: 20 μm). (B) Images of MMP-1-DsRed transfected live cells. The data show that all fluorescent signal (green, red, blue,) demonstrate partial co-localization (inset), with white pixels where all three is simultaneously present (bars 20 μm).
Mentions: General paradigm is that MMP-1 secreted and deposited in the extracellular matrix as an inactive zymogen by various cell types where it is subsequently activated [47]. To investigate further if MMP-1 cellular production and localization can overlap with the underlying collagen matrix degradation beneath the cells, live cell fluorescent imaging in combination with DQ-collagen, MMP1-DsRed fusion protein expression and MMP-1 cleavage specific Fluorogenic Substrate-III (Fig. 4A and B; Refs. [48, 49]) were used in invadopodia assays.

Bottom Line: Inhibition of prenylation by farnesyl transferase inhibitor (FTI-276) decreased extracellular MMP-1 and subsequently reduced invasiveness by 30%.Parallel, stable cell line RNAi knockdown of MMP-1 confirmed its role in cellular invasiveness.These results indicate that MMP-1 directional delivery to podia structures is involved in the invasive activity of sarcoma cells, and this process is prenylation sensitive.

View Article: PubMed Central - PubMed

Affiliation: Sarcoma Biology Laboratory of Sylvester Comprehensive Cancer Center, University of Miami, Miller School of Medicine, FL, USA. ngaramszegi@med.miami.edu

Show MeSH
Related in: MedlinePlus