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J Biol Chem, Vol. 274, Issue 36, 25576-25582, September 3, 1999

Transcriptional Regulation of the Platelet-derived Growth Factor alpha  Receptor Gene via CCAAT/Enhancer-binding Protein-delta in Vascular Smooth Muscle Cells

Tomikazu Fukuoka, Yutaka Kitami, Takafumi Okura, and Kunio Hiwada

From the Second Department of Internal Medicine, Ehime University School of Medicine, Onsen-gun, Ehime 791-0295, Japan

Inflammatory cytokines stimulate the proliferation of vascular smooth muscle cells (VSMC) and play a pivotal role in the pathogenesis of vascular diseases including atherosclerosis and restenosis. Mitogenic response of interleukin-1beta (IL-1beta ) on VSMC is thought to be mediated by induction of endogenous platelet-derived growth factor (PDGF), especially PDGF-AA. Although the action of PDGF-AA is mediated by its specific receptor, PDGFalpha -receptor (PDGFalpha R), very little is known about the regulatory mechanism of PDGFalpha R gene expression in VSMC. To understand the mechanism, we studied the transcriptional control of the PDGFalpha R gene in VSMC after treatment with IL-1beta . IL-1beta (10 ng/ml) drastically increased both PDGFalpha R and CCAAT/enhancer-binding protein delta  (C/EBPdelta ) mRNA levels in a time dependent manner. A rapid induction of C/EBPdelta mRNA within 30 min was followed by slower emergence of PDGFalpha R mRNA, which reached the maximum level in 12 h, whereas C/EBPdelta mRNA was detectable at 30 min and reached the maximum level at 3 h. Electromobility shift and supershift assays revealed that IL-1beta markedly increased DNA-protein complex, which was mainly composed of C/EBPbeta and/or -delta . Both Western blotting and immunohistochemistry demonstrated that either C/EBPbeta or -delta expression was induced by IL-1beta exclusively in nuclei of VSMC. On the other hand, overexpression of C/EBPdelta specifically transactivated the promoter activity of the PDGFalpha R gene and significantly enhanced VSMC proliferation in PDGF-treated cells. We conclude that induction of PDGFalpha R expression is mainly mediated by C/EBPdelta expression in VSMC, and a high level of C/EBPdelta expression may be involved in the pathogenesis of atherosclerosis and restenosis.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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