Agonist-induced down-regulation of alpha(1B)-adrenergic receptor in HEK293 cells transfected with alpha(1B) cDNA.
HEK293 cells stably expressing hamster alpha(1B)-adrenergic receptor (alpha(1B)-AR) were used to observe the effect of nonepinephrine (NE) on alpha(1B)-AR gene expression. Radioligand binding assys and RNase protection assays were used to determine alpha(1B)-AR number and the mRNA level, respectively. Exposure (2-24 h) of HEK293 cella to NE (10 mumol) caused a decrease in alpha(1B)-AR mRNA with maximum change found at the 4th hour. and in alpha(1B)-AR density at the 24th hour. NE-induced decrease in alpha(1B)-AR mRNA was inhibited by protein kinase C (PKC) inhibitor calphostin C (0.1. mumol) and mimicked by PKC activator PMA (1 mumol). Nuclear run-off transcription assay showed that treatment of the cells with NE (10 mumol) exerted no effect on the transcription rate of alpha(1B)-AR, After the synthesis of new RNAs was inhibited by actinomycin D, NE muld not accelerate the degradation of alpha(1B)-AR mRNA. The results suggested that in the HEK293 cells NE muld induce the down-regulation of alpha(1B)-AR, and the effects were mediated by PKC pathway. NE could not alter the transcription rate of alpha(1B)-AR mRNA, but it might induce the synthesis of some factors and indirectly accelerate the degradation.
Kaiming
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Institute of Vascular Medicine, Third Hospital, Beijing Medical University, 100083, Beijing, China.
Qide
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Bin
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Youyi
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