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首页> 外文期刊>Journal of bone and mineral research: the official journal of the American Society for Bone and Mineral Research >Involvement of CCAAT enhancer binding protein transcription factors in the regulation of prostaglandin G/H synthase 2 expression by interleukin-1 in osteoblastic MC3T3-E1 cells.
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Involvement of CCAAT enhancer binding protein transcription factors in the regulation of prostaglandin G/H synthase 2 expression by interleukin-1 in osteoblastic MC3T3-E1 cells.

机译:CCAAT增强子结合蛋白转录因子参与成骨细胞MC3T3-E1细胞中白介素1调节前列腺素G / H合酶2表达的作用。

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摘要

Interleukin-1 (IL-1) stimulates prostaglandin production in bone by a rapid and transient activation of prostaglandin G/H synthase 2 (PGHS-2) gene expression. In osteoblastic MC3T3-E1 cells, IL-1 caused a transient increase in PGHS-2 messenger RNA (mRNA), which peaked at 2 h. IL-1 caused a 2- to 4-fold activation of a 371-base pair (bp) murine PGHS-2 promoter/luciferase construct in stable transfectants. This response mapped to a proximal promoter element(s) located between -150 and -40 bp. This region contains a putative CCAAT enhancer binding protein (C/EBP) site (centered at -135 bp), which shows enhanced binding of C/EBPbeta and C/EBPdelta by mobility shift analysis after IL-1 treatment. A transient cotransfection approach was used to examine the effects of C/EBPbeta and C/EBPdelta overexpression. IL-1 caused a maximal 3- to 7-fold stimulation of PGHS-2 promoter activity after 2.5 h. Overexpression of murine C/EBPbeta and C/EBPdelta caused a dose-dependent increase in basal and IL-1-stimulated luciferase activity. C/EBPdelta caused a greater enhancement of basal and IL-1-stimulated promoter activity than C/EBPbeta, suggesting that C/EBPdelta is a stronger transactivator. Overexpression of p20C/EBPbeta, a dominant negative inhibitor of C/EBP function, blocked the stimulation of PGHS-2 promoter activity by IL-1 and blocked the ability of overexpressed C/EBPbeta and C/EBPdelta to increase basal and IL-1-stimulated promoter activity. Mutagenesis of the C/EBP site reduced, but did not abolish, the stimulation of PGHS-2 promoter activity by IL-1 and blunted the effect of overexpressed C/EBPdelta on basal and IL-1-stimulated promoter activity. These results suggest an essential role for C/EBPbeta and C/EBPdelta in the induction of PGHS-2 gene expression by IL-1 in osteoblastic cells.
机译:白介素-1(IL-1)通过快速和短暂激活前列腺素G / H合酶2(PGHS-2)基因表达来刺激骨骼中前列腺素的产生。在成骨细胞MC3T3-E1细胞中,IL-1引起PGHS-2信使RNA(mRNA)的短暂增加,在2小时达到峰值。 IL-1在稳定的转染子中引起371个碱基对(bp)的鼠PGHS-2启动子/荧光素酶构建体的2到4倍活化。该应答映射到位于-150和-40bp之间的近端启动子元件。该区域包含推定的CCAAT增强子结合蛋白(C / EBP)位点(中心位于-135 bp),通过IL-1处理后的迁移率变化分析显示C / EBPbeta和C / EBPdelta的结合增强。瞬时共转染方法用于检查C / EBPbeta和C / EBPdelta过表达的影响。 2.5小时后,IL-1对PGHS-2启动子活性产生了最大的3至7倍刺激。鼠C / EBPbeta和C / EBPdelta的过表达引起基础和IL-1刺激的荧光素酶活性的剂量依赖性增加。 C / EBPdelta引起的基础和IL-1刺激的启动子活性比C / EBPbeta更大,表明C / EBPdelta是更强的反式激活剂。 p20C / EBPbeta(C / EBP功能的主要负性抑制剂)的过表达,阻断了IL-1对PGHS-2启动子活性的刺激,并阻断了过表达的C / EBPbeta和C / EBPdelta增加基础和IL-1的能力。刺激的启动子活性。 C / EBP位点的诱变减少但没有消除IL-1对PGHS-2启动子活性的刺激,并减弱了过度表达的C / EBPdelta对基础和IL-1刺激的启动子活性的影响。这些结果表明C / EBPbeta和C / EBPdelta在成骨细胞中IL-1诱导PGHS-2基因表达中起着至关重要的作用。

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