Woeller Collynn F, Flores E'Lissa, Pollock Stephen J, Phipps Richard P
Department of Environmental Medicine, School of Medicine and Dentistry, University of Rochester, Rochester, New York, USA.
Toxicol Sci. 2017 Jun 1;157(2):305-319. doi: 10.1093/toxsci/kfx046.
Environmental chemicals termed "obesogens" disrupt the endocrine system to promote adipogenesis and obesity. Tetrabromobisphenol-A (TBBPA) has been reported to increase adipogenesis; however, the mechanism(s) of action are unclear. Thy1 (CD90) is a glycophosphatidylinositol-anchored membrane protein that serves as a marker for stem cells and also plays an important role in regulating adipogenesis and obesity. We investigated whether or not TBBPA promotes adipogenesis in human and mouse cells by reducing Thy1 levels. We further sought to identify the molecular mechanism(s) whereby TBBPA targets Thy1 expression. Mouse and human cells were exposed to TBBPA, and Thy1 expression was analyzed using flow cytometry, Western blotting, and qPCR. We tested whether microRNAs predicted to target Thy1 (miR-103 and miR-107) were upregulated by TBBPA using quantitative PCR assays. We also determined if Thy1 mRNA was a bona fide miR-103/107 target. Our results show that Thy1 expression was reduced in both human and mouse cells after exposure to TBBPA. Both Thy1 mRNA and protein levels were decreased by low-dose TBBPA exposure. TBBPA reduced Thy1 levels and further increased adipogenesis when an adipogenic medium was used. Mechanistically, we show that miR-103 and miR-107 are induced by TBBPA and that miR-103 targets Thy1 to reduce its expression. Our results reveal for the first time that Thy1 is a target of TBBPA. Furthermore, our data support the concept that Thy1 is a key marker targeted by environmental chemicals that promote adipogenesis and obesity.
被称为“致肥胖因子”的环境化学物质会扰乱内分泌系统,从而促进脂肪生成和肥胖。据报道,四溴双酚A(TBBPA)可增加脂肪生成;然而,其作用机制尚不清楚。Thy1(CD90)是一种糖基磷脂酰肌醇锚定膜蛋白,作为干细胞的标志物,在调节脂肪生成和肥胖中也起着重要作用。我们研究了TBBPA是否通过降低Thy1水平来促进人和小鼠细胞的脂肪生成。我们进一步试图确定TBBPA靶向Thy1表达的分子机制。将小鼠和人类细胞暴露于TBBPA,并使用流式细胞术、蛋白质免疫印迹和定量聚合酶链反应(qPCR)分析Thy1表达。我们使用定量PCR测定法测试了预测靶向Thy1的微小RNA(miR-103和miR-107)是否被TBBPA上调。我们还确定了Thy1信使核糖核酸(mRNA)是否是miR-103/107的真正靶点。我们的结果表明,人和小鼠细胞在暴露于TBBPA后Thy1表达均降低。低剂量TBBPA暴露可降低Thy1 mRNA和蛋白质水平。当使用成脂培养基时,TBBPA降低了Thy1水平并进一步增加了脂肪生成。从机制上讲,我们表明miR-103和miR-107由TBBPA诱导,并且miR-103靶向Thy1以降低其表达。我们的结果首次揭示Thy1是TBBPA的一个靶点。此外,我们的数据支持这样一种概念,即Thy1是促进脂肪生成和肥胖的环境化学物质所靶向的关键标志物。