Endothelin-1 causes a prolonged protein kinase C activation and acts as a co-mitogen in vascular smooth muscle cells

Abstract

Endothelin-1 (ET-1) is known to act via G-protein coupled receptors. It has therefore been suggested that any mitogenic activity it may possess, is due to activation of phospholipase C and protein kinase C (PKC). We have therefore examined both the ability of ET-1 to act as a mitogen and its ability to activate PKC. We found that ET-1 significantly increased thymidine incorporation and enhanced platelet-derived growth factor-induced DNA synthesis, as well as causing a prolonged translocation of PKC to the cell membrane. ET-1 significantly increased PKC dependent phosphorylation of two specific substrates. The phosphorylation of MBP4-14 (from myelin basic protein) was partially dependent on extracellular Ca2+, implicating activation of PKC-alpha, whereas phosphorylation of the so called epsilon-peptide was Ca(2+)-independent and prolonged. This could be due either to the delta or zeta isoform of PKC, known to be present in these cells. However, ET-1 induced little proliferation of PKC activity in a transformed smooth muscle cell line, DDT1 MF-2, which lacks expression of the PKC-alpha isoform, but expresses the zeta-isoform. Thus, it would appear the ET-1-induced mitogenicity in smooth muscle cells may be related to the sustained, Ca(2+)-independent activation of PKC-delta.