Requirement for protein kinase C θ for cell cycle progression and formation of actin stress fibers and filopodia in vascular endothelial cells

Abstract

Activation of the protein kinase C (PKC) family with phorbol esters induces endothelial proliferation and angiogenesis, but which of the events that constitute angiogenesis are affected by individual members of the PKC family is unknown. In rat capillary endothelial (RCE) cells, serum stimulation increased expression of a single PKC isoenzyme, PKCθ, and its translocation to the periphery. Conditional overexpression of a dominant- negative mutant of PKCθ markedly inhibited RCE proliferation, as well as closure of a 'wound' by RCE migration and formation of capillary rings and tubules in vitro. PKCθ inhibition delayed the endothelial cell cycle at the Germ phase and prevented formation of actin stress fibers and filopodia but not lamellipodia. The defect in cell morphology and wound closure in PKCθ- kn cells was reversed by overexpressing kinase-active PKCθ, indicating that these RCE functions depend upon PKCθ substrates. Thus, PKCθ is required for multiple processes essential for angiogenesis and wound repair, including endothelial mitosis, maintenance of a normal actin cytoskeleton, and formation of an enclosed tube.