Institute of Cellular Biology and Pathology "N. Simionescu", 050568 Bucharest, Romania.
Laboratory of Biochemistry, University of Crete Medical School and Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology of Hellas, Heraklion, 71003 Crete, Greece.
Int J Mol Sci. 2019 Mar 23;20(6):1471. doi: 10.3390/ijms20061471.
Apolipoprotein E (apoE) is mainly secreted by hepatocytes and incorporated into most plasma lipoproteins. Macrophages, which accumulate cholesterol and are critical for the development of the atherosclerotic plaque, are also an important, albeit smaller, apoE source. Distal regulatory elements control cell-specific activity of the apoE promoter: multienhancers (ME.1/2) in macrophages and hepatic control regions (HCR-1/2) in hepatocytes. A member of AP-1 cell growth regulator, c-Jun regulates the transcription of various apolipoproteins and proinflammatory molecules implicated in atherosclerosis. We aimed to investigate the effect of c-Jun on apoE expression in macrophages versus hepatocytes and to reveal the underlying molecular mechanisms. Herein we show that c-Jun had an opposite, cell-specific effect on apoE expression: downregulation in macrophages but upregulation in hepatocytes. Transient transfections using ME.2 deletion mutants and DNA pull-down (DNAP) assays showed that the inhibitory effect of c-Jun on the apoE promoter in macrophages was mediated by a functional c-Jun binding site located at 301/311 on ME.2. In hepatocytes, c-Jun overexpression strongly increased apoE expression, and this effect was due to c-Jun binding at the canonical site located at -94/-84 on the apoE proximal promoter, identified by transient transfections using apoE deletion mutants, DNAP, and chromatin immunoprecipitation assays. Overall, the dual effect of c-Jun on apoE gene expression led to decreased cholesterol efflux in macrophages resident in the atherosclerotic plaque synergized with an increased level of systemic apoE secreted by the liver to exacerbate atherogenesis.
载脂蛋白 E(apoE)主要由肝细胞分泌,并整合到大多数血浆脂蛋白中。巨噬细胞是胆固醇的主要蓄积细胞,对动脉粥样硬化斑块的形成至关重要,也是 apoE 的重要来源,尽管其含量较少。远端调节元件控制 apoE 启动子的细胞特异性活性:巨噬细胞中的多增强子(ME.1/2)和肝细胞中的肝控制区(HCR-1/2)。AP-1 细胞生长调节剂的一个成员,c-Jun 调节各种与动脉粥样硬化有关的载脂蛋白和前炎症分子的转录。我们旨在研究 c-Jun 对巨噬细胞与肝细胞中 apoE 表达的影响,并揭示潜在的分子机制。在此,我们表明 c-Jun 对 apoE 表达具有相反的细胞特异性作用:在巨噬细胞中下调,而在肝细胞中上调。使用 ME.2 缺失突变体的瞬时转染和 DNA 拉下(DNAP)测定表明,c-Jun 对巨噬细胞中 apoE 启动子的抑制作用是由位于 ME.2 上 301/311 的功能性 c-Jun 结合位点介导的。在肝细胞中,c-Jun 的过表达强烈增加了 apoE 的表达,这种效应归因于 c-Jun 在 apoE 近端启动子上位于-94/-84 的典型结合位点的结合,该位点通过使用 apoE 缺失突变体、DNAP 和染色质免疫沉淀测定的瞬时转染来鉴定。总的来说,c-Jun 对 apoE 基因表达的双重作用导致动脉粥样硬化斑块中驻留的巨噬细胞胆固醇流出减少,与肝脏分泌的全身性 apoE 水平增加协同作用,加剧了动脉粥样硬化的发生。