Increase of urokinase-type plasminogen activator receptor expression in human gingival fibroblasts by Porphyromonas gingivalis lipopolysaccharide.
BACKGROUND
Urokinase-type plasminogen activator receptor (uPAR) presenting on the cell surface with glycosylphosphatidylinositol (GPI) anchor is a key component in the plasminogen activator (PA)-plasmin system and plays a critical role in extracellular matrix degradation. It is believed that uPAR serves to localize and accelerate this generation system. In this study, we examined the levels of both uPA and uPAR in human gingival fibroblasts treated with Porphyromonas gingivalis lipopolysaccharide (LPS).


METHODS
Human gingival fibroblasts from the seventh to tenth doubling passages were plated at 5x10(4) cells per well in 24-well plates. The confluent-stage cells were cultured for 24 hours in alpha-MEM medium containing 2% fetal calf serum, after which they were incubated with P. gingivalis LPS. PA activity was measured using plasminogen and plasmin substrate S2251.


RESULTS
PA activity in the cell lysate was increased by LPS and reached maximum at 1 microg/ml LPS after being incubated for 8 hours. PA activity released by phosphatidylinositol-specific phospholipase C, which detaches the GPI anchor, was also increased by LPS. The activity was inhibited by amiloride, which is a specific inhibitor for uPA. LPS increased the protein and mRNA levels of both uPA and uPAR in gingival fibroblasts.


CONCLUSIONS
These findings suggest that increase of the uPAR level by LPS may play an important role in the progression of periodontal diseases through pericellular proteolysis.
D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000201:enzymology; D005347:Fibroblasts / Q000201:enzymology* / Q000382:microbiology; D005881:Gingiva / Q000166:cytology / Q000201:enzymology* / Q000382:microbiology; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism*; D010510:Periodontal Diseases / Q000201:enzymology / Q000382:microbiology; D010960:Plasminogen Activators / Q000096:biosynthesis*; D016966:Porphyromonas gingivalis / Q000737:chemistry / Q000472:pathogenicity*; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D055293:Receptors, Urokinase Plasminogen Activator; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis*