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Impact of rosiglitazone on the expression of β3-AR in the stable cell lines expressed β3-AR gene
Published/Copyright:
October 31, 2007
Abstract
Background: The aim of the present study was to investigate the effect of rosiglitazone, a peroxisome proliferator-activated receptor γ2 (PPAR-γ2) agonist, on the expression of β3-adrenergic receptor (β3-AR) at transcriptional and translational level.
Methods: We cloned the cDNA sequences of human PPAR-γ2 (hPPAR-γ2) gene and human wild type and mutant β3-adrenergic receptor (hβ3-AR) genes and established their eukaryotic expression vectors. The pcDNA3.1/hβ3-AR (mutant and wild type) was transfected into SH-SY5Y cells using electroporation method. The expression level of β3-AR protein was determined by Western blot analysis.
Results: Our results showed that the reverse transcription-PCR products were consistent with theoretical fragment sizes of human PPAR-γ2 (1544 bp) and human β3-AR genes (1578 bp). The sequence analysis of PPAR-γ2 and β3-AR genes showed that the fragment sizes were the same as that of human PPAR-γ2 and human β3-AR genes in Genebank. The pcDNA3.1/hβ3-AR (mutant and wild type) was successfully cloned to SH-SY5Y cells. We found that the expression of β3-AR protein was significantly inhibited by rosiglitazone in a concentration-dependent manner in SH-SY5Y cell lines stably expressed β3-AR genes.
Conclusions: The results suggest that rosiglitazone has a concentration-dependent inhibitory effect on the expression of β3-AR protein, and this inhibitory effect may be due to activation of PPAR-γ2 receptor.
Clin Chem Lab Med 2007;45:1511–6.
Keywords: β3-adrenergic receptor; peroxisome proliferator-activated receptor γ2; rosiglitazone; stable cell lines; transfection
Received: 2007-5-10
Accepted: 2007-7-16
Published Online: 2007-10-31
Published in Print: 2007-11-01
©2007 by Walter de Gruyter Berlin New York
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Keywords for this article
β3-adrenergic receptor;
peroxisome proliferator-activated receptor γ2;
rosiglitazone;
stable cell lines;
transfection
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