HUMAN PLACENTAL ENDOTHELIN - EXPRESSION OF ENDOTHELIN-1 MESSENGER-RNA BY HUMAN PLACENTAL FIBROBLASTS IN CULTURE

Endothelin-1 (ET-L) mRNA is expressed by the human placenta in a developmentally regulated manner and has been shown to stimulate the growth of placental mesenchymal cells. The ability of placental fibroblasts to express preproET-1 mRNA was studied to determine if ET-1 could potentially participate via autocrine mechanisms in the proliferation of placental fibroblasts. Fibroblasts were isolated from normal placentae at various gestational ages (7-19 weeks and term) and their abilities to express preproET-1 mRNA in culture evaluated by Northern analysis. Sparse, rapidly growing cultures of placental fibroblasts expressed preproET-1 mRNA at each gestational age in the presence of 10% FBS. The regulation of preproET-1 expression in placental fibroblasts was studied by exposing cells to known mitogenic stimuli. Quiescent, confluent monolayers of placental fibroblasts expressed no detectable levels of preproET-1 mRNA under basal conditions. Epidermal growth factor (EGF, 10 ng/ml), transforming growth factor-beta 1 (TGF-beta 1, 5 ng/ml), or interleukin 1 beta (IL-1 beta) alone, had no significant effect on steady state preproET-1 mRNA levels. Cycloheximide, an inhibitor of protein synthesis, increased the steady state levels of preproET-1 mRNA at a concentration of 10 mu g/ml. In the presence cycloheximide, IL-1 beta markedly stimulated preproET-1 mRNA expression, whereas EGF was less effective. TGF-beta 1 had no effect in the presence or absence of cycloheximide. In contrast, 12-O-tetradecanoylphorbol 13-acetate (TPA, 20 nM) exerted a small stimulatory effect on preproET-1 mRNA expression which was not influenced by cycloheximide. These data demonstrate that placental fibroblasts express preproET-1 mRNA while actively growing and that its expression is regulated by specific growth promoting stimuli. These data suggest that placental ET-1 may act, in part, via autocrine mechanisms to regulate the growth or function of placental fibroblasts.