Urokinase and tissue-type plasminogen activator stimulate human vascular smooth muscle cell migration

The objective of this study was to investigate the role of the plasminogen activation system in the migration of human vascular smooth muscle cells in vitro. After wounding of confluent human smooth muscle cell cultures by stripping cells from their extracellular matrix, cells start to migrate from the wounded edge into the denuded area. Addition of plasmin to the culture medium resulted in an approximately 50% increase of migrated cells after 24 hours. The plasmin inhibitor aprotinin was able to reduce this effect to control levels. Migration could also be stimulated by addition of urokinase-type plasminogen activator (HMW-u-PA) (30%) or tissue-type plasminogen activator (t-PA) (28%). Simultaneous addition of aprotinin reduced this increase below control levels, indicating that both HMW-u-PA and t-PA mediated plasminogen activation contributes to smooth muscle cell migration. Addition of low molecular weight u-PA (LMW-u-PA), a u-PA form lacking the receptor binding domain, or the aminoterminal fragment of u-PA (ATF), lacking the active site, had no effect on migration. These results suggest that both t-PA and u-PA can contribute to human smooth muscle cell migration in vitro, most likely via plasminogen activation. For the stimulation of migration by u-PA, activity as well as binding to its cell-surface receptor appears to be involved.