Abstract

The effects of epidermal growth factor, basic fibroblast growth factor, and platelet-derived growth factor-BB on angiotensin type 1 (AT1) receptor gene expression were examined in rat thoracic aorta vascular smooth muscle cells (VSMC) in culture. Incubation of serum-deprived VSMC with 20 ng/ml epidermal growth factor, 20 ng/ml basic fibroblast growth factor, or 50 ng/ml platelet-derived growth factor-BB reduces AT1 receptor mRNA levels, as assessed by Northern hybridization analysis, to approximately 30% of control levels. This effect is maximal 4 hr after addition of each growth factor to the culture medium and is sustained for up to 24 hr of incubation after a single dose. There is a correlative loss of membrane-associated AT1 receptors and angiotensin II-stimulated inositol phosphate production after 24 hr of growth factor treatment. The half-life of AT1 receptor mRNA is reduced significantly by growth factors, compared with that for cells treated with actinomycin D alone to block transcription. This suggests that growth factors activate a mechanism that involves post-transcriptional destabilization of AT1 receptor mRNA. This effect can be blocked by prior treatment of VSMC with actinomycin D or cycloheximide, suggesting that the effect of the growth factors on AT1 receptor gene expression is mediated through induction of an unknown gene or genes that function to destabilize AT1 receptor mRNA and that mRNA translation is essential for the destabilizing effect. Nuclear run-on assays reveal that the growth factors also significantly reduce the rate of de novo AT1 receptor gene transcription. Thus, down-regulation of AT1 receptor gene expression by growth factors also appears to involve mechanisms that decrease the rate of AT1 receptor gene transcription. These data reveal marked down-regulation of AT1 receptor gene expression in VSMC by growth factor receptor activation, through mechanisms that involve both attenuation of transcription and post-transcriptional mRNA destabilization.