Inhibition of tumor invasion by genomic down-regulation of matriptase through suppression of activation of receptor-bound pro-urokinase.
Urokinase-type plasminogen activator (uPA) degrades the extracellular matrix and plays critical roles in tumor invasion and metastasis. Matriptase, a membrane-bound serine protease, was shown to activate uPA in a uPA receptor-free, solution-based study. We now investigate whether matriptase affects activation of receptor-bound uPA and contributes to the invasiveness of HRA human ovarian cancer cells in vitro and tumor behavior in nude mice. Here we show the following. 1) uPA expression was effectively stimulated by TGF-beta1 in HRA cells. 2) Antisense (AS)-matriptase transfection achieved a marked inhibition of receptor-bound pro-uPA activation without altering expression of uPA and uPA receptor mRNA and proteins, irrespective of whether cells were stimulated with TGF-beta1. 3) Tumor cell receptor-bound pro-uPA could be efficiently cleaved by matriptase to generate enzymatically active two-chain uPA. Thus, matriptase can substitute for plasmin in the proteolytic activation of pro-uPA to enzymatically active uPA. 4) The AS-matriptase-treated cells had a decreased ability to invade an extracellular matrix layer, as compared with control cells. 5) When the AS-matriptase-treated cells were injected intraperitoneally into nude mice, the mice developed smaller tumors. Our data identify a novel role for matriptase for activation of receptor-bound uPA.
Suzuki
Mika
M
Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Handayama 1-20-1, Hamamatsu, Shizuoka 431-3192, Japan.
Kobayashi
Hiroshi
H
Kanayama
Naohiro
N
Saga
Yasushi
Y
Suzuki
Mitsuaki
M
Lin
Chen-Yong
CY
Dickson
Robert B
RB
Terao
Toshihiko
T
D000818:Animals; D001667:Binding, Competitive; D015152:Blotting, Northern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D004396:Coloring Agents / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005109:Extracellular Matrix / Q000201:enzymology / Q000378:metabolism; D005341:Fibrinolysin / Q000378:metabolism; D005434:Flow Cytometry; D016678:Genome; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D051379:Mice; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010534:Peritoneal Neoplasms / Q000473:pathology; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012697:Serine Endopeptidases; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism; D053773:Transforming Growth Factor beta1; D014360:Trypsin Inhibitor, Kunitz Soybean / Q000096:biosynthesis; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis*
D004396:Coloring Agents; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D016376:Oligonucleotides, Antisense; C524898:PLAUR protein, human; C524899:Plaur protein, mouse; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; C109148:SPINT2 protein, human; C508960:TGFB1 protein, human; D013778:Tetrazolium Salts; C508961:Tgfb1 protein, mouse; D013844:Thiazoles; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D014360:Trypsin Inhibitor, Kunitz Soybean; D012697:Serine Endopeptidases; C485595:matriptase; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; C022616:thiazolyl blue; C067551:saruplase