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Neutrophil-derived serine proteinases enhance membrane type-1 matrix metalloproteinase-dependent tumor cell invasion
P Shamamian1, B J Pocock, J D Schwartz
1Department of Surgery, S. Arthur Localio Laboratory for Surgical Research, Kaplan Comprehensive Cancer Center, New York, NY, USA.
Background:
Matrix metalloproteinase-2 degrades a variety of basement membrane components and is essential for tumor invasion. We have previously reported that membrane type-1 matrix metalloproteinase (MT1-MMP) cooperates with neutrophil-derived serine proteinases (NDPs; elastase, cathepsin G, protease-3) to activate matrix metalloproteinase-2. We therefore hypothesized that NDPs enhance tumor-cell invasion.
Methods:
Clones of human HT1080 fibrosarcoma cells transfected with MT1-MMP sense (HT-SE) or antisense CDNA (HT-AS) were used. These cells express either high (HT-SE) or extremely low levels (HT-AS) of MT1-MMP relative to nontransfected HT1080 cells (HT-WT). The cells were incubated in the presence or absence of purified NDP, with or without alpha 1-antitrypsin or the MMP inhibitor batimastat. Cell invasion was measured with the use of Boyden chambers with polycarbonate membranes coated with a reconstituted extracellular matrix.
Results:
Under control conditions HT-WT and HT-SE cells were 4-fold more invasive than HT-AS cells. The addition of NDP increased HT-WT and HT-SE cell invasion 60% to 100% but had no effect on HT-AS cells. alpha 1-antitrypsin or batimastat did not decrease the baseline invasiveness of HT-WT and HT-SE cells; however, they abrogated the stimulatory effect of NDP.
Conclusions:
HT1080 cell invasion depends on MT1-MMP expression. MT1-MMP overexpression does not increase invasiveness by itself. NDPs increase invasion by MT1-MMP expressing cells by activating matrix metalloproteinase-2.
Insights
Neutrophil-derived proteinases (NDPs) enhance tumor cell invasion by activating matrix metalloproteinase-2 (MMP-2). This effect requires membrane type-1 matrix metalloproteinase (MT1-MMP) expression, as NDPs stimulate invasion only in MT1-MMP-expressing cells.
Area of Science:
- Oncology
- Biochemistry
- Cell Biology
Background:
- Matrix metalloproteinase-2 (MMP-2) is crucial for tumor invasion due to its role in degrading basement membrane components.
- Membrane type-1 matrix metalloproteinase (MT1-MMP) collaborates with neutrophil-derived proteinases (NDPs) to activate MMP-2.
- This interaction suggests NDPs may promote tumor cell invasion.
Purpose of the Study:
- To investigate the role of neutrophil-derived proteinases (NDPs) in enhancing tumor cell invasion.
- To determine if NDPs stimulate invasion through matrix metalloproteinase-2 (MMP-2) activation.
- To elucidate the dependence of this process on membrane type-1 matrix metalloproteinase (MT1-MMP) expression.
Main Methods:
- Human fibrosarcoma HT1080 cells with varying levels of MT1-MMP expression (high, low, or wild-type) were utilized.
- Cells were treated with purified NDPs, with or without inhibitors (alpha 1-antitrypsin or batimastat).
- Cell invasion was quantified using Boyden chambers with a reconstituted extracellular matrix.
Main Results:
- Cells expressing MT1-MMP showed significantly higher invasion compared to those with low MT1-MMP levels.
- NDP addition increased invasion in MT1-MMP-expressing cells by 60-100% but had no effect on low-expression cells.
- Inhibitors blocked the stimulatory effect of NDPs, indicating MMP-2 activation is key.
Conclusions:
- Tumor cell invasion is dependent on MT1-MMP expression.
- MT1-MMP overexpression alone does not drive increased invasiveness.
- NDPs enhance invasion in MT1-MMP-expressing cells by activating MMP-2, highlighting a novel pathway in tumor metastasis.