Vasoactive intestinal peptide and epidermal growth factor: co-mitogens or inhibitors of keratinocyte proliferation in vitro?

Abstract

Vasoactive intestinal peptide (VIP) is a neuropeptide with a broad range of biological activities in various tissues. Interactions of VIP and epidermal growth factor (EGF) are of particular interest for dermatology. They may be either co-mitogenic or inhibitory. HaCaT keratinocytes cultivated under serum-free conditions in vitro have been used to investigate the interactions of VIP and EGF. EGF was found to induce cell growth, whereas preincubation with VIP inhibited EGF-induced proliferation in a dose-dependent manner. Maximum growth inhibition was 46% (p < 0.01) at a VIP concentration of 10(-7) M. EGF-induced growth is mediated by tyrosine kinase (TK). Therefore we studied the effect of VIP on TK activity. Cells were incubated with VIP (10(-13)-10(-7) M) for 48 h and stimulated with EGF at a final concentration of 500 ng/ml. SDS-PAGE and Western blot with the antibody RC20H against TK were performed. We found a dose dependent decrease of EGF receptor TK activity. At VIP concentration of 10(-7) M a residual TK activity of 65% was detected. To investigate the possibly involved signal transduction pathways, we performed inhibition experiments with wortmannin, pertussis toxin, 2'5'diacylglycerol and adenosine-3':5'-mono-phosphorothioate. However, none of the inhibitors was effective in abolishing growth inhibition by VIP. VIP was shown to be growth inhibitory for human keratinocytes. The data suggest that EGF receptor TK is involved in signal transduction of VIP. Thus TK activity is a possible common target of both EGF- and VIP-induced cellular responses.