Oxidized LDL inhibits vascular endothelial cell morphogenesis in culture

Human umbilical cord vein endothelial cells can be induced to undergo morphogenesis (tube formation) by phorbol ester (TPA) when cultured on or in three-dimensional collagen gels. Induction of morphogenesis by TPA is accompanied by increased activity of the collagenase gene transcription factors, ETS1 and API, and the elaboration of collagenase by the endothelial cells. In the present study, we used endothelial cell elongation as a measure of morphogenesis and showed that oxidized low density lipoprotein (oxLDL) inhibited endothelial cell migration in monolayer cultures and TPA-induced morphogenesis in collagen gels in a dose-dependent manner. Moreover, the inhibition was positively correlated with the extent of LDL oxidation. In contrast, native LDL stimulated cell migration and TPA-induced morphogenesis under the same culture conditions. However, in the absence of TPA, LDL showed no effect on EC morphogenesis. Further studies showed that inhibition of TPA-induced endothelial cell morphogenesis by oxLDL is correlated with suppression of the protein kinase C (PKC) and ETS1/AP1 activities. The results indicated that the inhibition of endothelial cell morphogenesis by oxLDL is probably mediated through inhibition of the TPA-activated PKC pathway and its subsequent suppression of the ETS1/AP1 activity. The results also indicated that EC migration can be mediated through PKC-dependent and independent pathways and only the former pathway can induce EC morphogenesis as well.