Urokinase upregulates matrix metalloproteinase-9 expression in THP-1 monocytes via gene transcription and protein synthesis.
Urokinase-type plasminogen activator (uPA) is suggested to exert its proliferatory, migratory and invasive action through binding with its membrane receptor, promoting pericellular proteolysis and mediating cell signal transduction. One of the possible actions of urokinase can be related to the regulation of activity and/or the expression of proteolytic enzymes participating in extracellular matrix degradation. In the present study, the role of uPA in regulating matrix metalloproteinase (MMP) expression and release by the monocyte cell line THP-1 was investigated. Recombinant uPA induced the release of MMP9/gelatinase B, as detected by zymography and Western blotting, and this release was abolished by actinomycin D and cycloheximide (inhibitors of DNA transcription and protein synthesis) and partially suppressed by monensin (an inhibitor of secretion). Proteolytically inactive urokinase with substitution of His(204) for Gln was able to reproduce about 70% of the effect induced by the wild-type recombinant uPA. The reverse transcription-PCR and Northern blot data indicated that the action of r-uPA on THP-1 cells resulted in formation of MMP9 mRNA, which depended on time, within 6-48 h, of the cell incubation with r-uPA. These results suggest that urokinase upregulates MMP9 expression in monocytes via MMP9 gene transcription and protein biosynthesis.
Menshikov
Mikhail
M
Institute of Experimental Cardiology, Cardiology Research Center, 3rd Cherepkovskaya St., 15a, 121552, Moscow, Russia. myumensh@cardio.ru
Elizarova
Eugenia
E
Plakida
Karina
K
Timofeeva
Angelika
A
Khaspekov
Georgy
G
Beabealashvilli
Robert
R
Bobik
Alex
A
Tkachuk
Vsevolod
V
D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D017931:DNA Primers; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics*; D009000:Monocytes / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*; D015854:Up-Regulation / Q000502:physiology*; D014568:Urokinase-Type Plasminogen Activator / Q000502:physiology*
D017931:DNA Primers; D012333:RNA, Messenger; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9