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Up-regulation of Axin2 by dexamethasone promotes adipocyte differentiation in ROB-C26 mesenchymal progenitor cells

机译:地塞米松上调Axin2促进ROB-C26间充质祖细胞中的脂肪细胞分化

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Dexamethasone (Dex) regulates osteoblastic and adipocytic differentiation in mesenchymal progenitor cells through regulation of Wnt/β-catenin signaling. To elucidate the regulatory mechanisms underlying the effects of Dex, we examine the expression of Axin2, which is an intracellular inhibitor of Wnt/β-catenin signaling, in ROB-C26 clonal mesenchymal progenitor cells (C26). We observed the induction of Axin2 mRNA in C26 cells in response to Dex treatment. Treatment with a glucocorticoid receptor (GR) antagonist, mifepristone, showed that Dex-induced up-regulation of Axin2 is mediated by the GR. In the absence of Dex, gene silencing by using Axin2-targeted short hairpin RNA increased the number of alkaline phosphatase (ALP)-positive and nuclear β-catenin-positive cells and ALP activity. In the presence of Dex, Axin2 knockdown resulted in an increased number of ALP-positive and nuclear β-catenin-positive cells. Furthermore, Axin2 knockdown in Dex-treated cells suppressed adipocyte differentiation (as determined by reduced Oil Red O staining), reduced the number of PPARγ-positive and aP2-positive cells and decreased the mRNA expression of PPARγ2 and aP2. These results suggest that Axin2 plays a key role in adipocyte and osteoblastic differentiation by controlling β-catenin expression.
机译:地塞米松(Dex)通过调节Wnt /β-catenin信号传导来调节间充质祖细胞中的成骨细胞和脂肪细胞分化。为了阐明Dex效应的潜在调控机制,我们检查了ROX-C26克隆间充质祖细胞(C26)中Axin2的表达,Axin2是Wnt /β-catenin信号的细胞内抑制剂。我们观察到在响应Dex处理的C26细胞中Axin2 mRNA的诱导。用糖皮质激素受体(GR)拮抗剂米非司酮治疗,表明Dex诱导的Axin2上调是由GR介导的。在没有Dex的情况下,使用Axin2靶向的短发夹RNA进行基因沉默会增加碱性磷酸酶(ALP)阳性和核β-catenin阳性细胞的数量以及ALP活性。在存在Dex的情况下,Axin2敲低导致ALP阳性和核β-catenin阳性细胞数量增加。此外,Dex处理的细胞中的Axin2敲低抑制了脂肪细胞的分化(通过减少的油红O染色确定),减少了PPARγ阳性和aP2阳性细胞的数量,并减少了PPARγ2和aP2的mRNA表达。这些结果表明Axin2通过控制β-catenin表达在脂肪细胞和成骨细胞分化中起关键作用。

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