HT-1080 fibrosarcoma cell matrix degradation and invasion are inhibited by the matrix-associated serine protease inhibitor TFPI-2/33 kDa MSPI.
The urokinase-urokinase receptor system plays a dominant role in the degradation and invasion of extracellular matrix (ECM) by tumor cells. In this system, urokinase bound to its cell receptor converts plasminogen to plasmin, a broad-spectrum serine protease that participates in the degradation and invasion of connective tissues by tumor cells. In this study, we evaluated whether these activities of plasmin are inhibited by a newly characterized human 32 kDa recombinant serine protease inhibitor called trypsin/tissue factor pathway inhibitor-2 (rTFPI-2). We found that rTFPI-2 dose-dependently inhibited fluid-phase plasmin as well as plasmin generated on the ECM and/or the cell surface of HT-1080 fibrosarcoma cells. The degradation of radiolabeled matrix as well as Matrigel invasion by these tumor cells is also inhibited by rTFPI-2 in a dose-dependent fashion. We have reported that rTFPI-2 is identical to 33 kDa extracellular matrix-associated serine protease inhibitor (33 kDa MSPI), whereas the 31 and 27 kDa MSPIs are under-glycosylated forms of the 33 kDa MSPI. We therefore evaluated the ability of MSPIs from the ECM of dermal fibroblasts to inhibit plasmin and found that the plasmin activity was effectively blocked by the MSPIs. We have also evaluated whether the HT-1080 cells synthesize and secrete the MSPIs and found that the synthesis and secretion of the MSPIs was undetectable in these cells. Collectively, our results suggest that rTFPI-2/33 kDa MSPI inhibits plasmin on the tumor cell and ECM surfaces as well as the degradation and invasion of matrix by HT-1080 fibrosarcoma cells.
Rao
C N
CN
Department of Dermatology, Northwestern University School of Medicine, Chicago, IL 60611-3008, USA.
Cook
B
B
Liu
Y
Y
Chilukuri
K
K
Stack
M S
MS
Foster
D C
DC
Kisiel
W
W
Woodley
D T
DT
D000970:Antineoplastic Agents / Q000494:pharmacology*; D003094:Collagen; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D005341:Fibrinolysin / Q000037:antagonists & inhibitors; D005347:Fibroblasts / Q000378:metabolism; D005354:Fibrosarcoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D006023:Glycoproteins / Q000494:pharmacology*; D006801:Humans; D007797:Laminin; D009361:Neoplasm Invasiveness; D011257:Pregnancy Proteins / Q000494:pharmacology*; D011509:Proteoglycans; D011994:Recombinant Proteins / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D012867:Skin / Q000166:cytology / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D004338:Drug Combinations; D006023:Glycoproteins; D007797:Laminin; D011257:Pregnancy Proteins; D011509:Proteoglycans; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; C021240:tissue-factor-pathway inhibitor 2; C059485:matrigel; D003094:Collagen; D005341:Fibrinolysin