Neutrophil-derived serine proteinases enhance membrane type-1 matrix metalloproteinase-dependent tumor cell invasion.
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.
Shamamian
P
P
Department of Surgery, S. Arthur Localio Laboratory for Surgical Research, Kaplan Comprehensive Cancer Center, New York, NY, USA.
Pocock
B J
BJ
Schwartz
J D
JD
Monea
S
S
Chuang
N
N
Whiting
D
D
Marcus
S G
SG
Galloway
A C
AC
Mignatti
P
P
D056649:Cathepsin G; D002403:Cathepsins / Q000502:physiology; D017077:Culture Media, Conditioned; D005354:Fibrosarcoma / Q000503:physiopathology; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000502:physiology*; D061965:Matrix Metalloproteinase Inhibitors; D053146:Myeloblastin; D009361:Neoplasm Invasiveness / Q000503:physiopathology*; D009504:Neutrophils / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000502:physiology; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000502:physiology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000502:physiology; D000515:alpha 1-Antitrypsin / Q000494:pharmacology
D017077:Culture Media, Conditioned; D061965:Matrix Metalloproteinase Inhibitors; D015842:Serine Proteinase Inhibitors; D013876:Thiophenes; D000515:alpha 1-Antitrypsin; D010649:Phenylalanine; C080985:batimastat; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D010196:Pancreatic Elastase; D053146:Myeloblastin; D020778:Matrix Metalloproteinase 2