Faculty of Pharmacy in Hradec Králové, Department of Biochemical Sciences, Charles University, Hradec Králové, Czech Republic; Norwegian Center for Stem Cell Research, Department of Immunology and Transfusion Medicine, Oslo University Hospital Rikshospitalet, Oslo, Norway.
Faculty of Pharmacy in Hradec Králové, Department of Biochemical Sciences, Charles University, Hradec Králové, Czech Republic.
Biochim Biophys Acta Gene Regul Mech. 2021 Oct;1864(10):194734. doi: 10.1016/j.bbagrm.2021.194734. Epub 2021 Jul 31.
Glutathione peroxidase 7 (GPx7) acts as an intracellular stress sensor/transmitter and plays an important role in adipocyte differentiation and the prevention of obesity related pathologies. For this reason, finding the regulatory mechanisms that control GPx7 expression is of great importance. As microRNAs (miRNAs) could participate in the regulation of GPx7 expression, we studied the inhibition of GPx7 expression by four selected miRNAs with relation to obesity and adipogenesis. The effect of the transfection of selected miRNAs mimics on GPx7 expression was tested in three cell models (HEK293, SW480, AT-MSC). The interaction of selected miRNAs with the 3'UTR of GPx7 was followed up on using a luciferase gene reporter assay. In addition, the levels of GPx7 and selected miRNAs in adipose tissue mesenchymal stem cells (AT-MSC) and mature adipocytes from four human donors were compared, with the changes in these levels during adipogenesis analyzed. Our results show for the first time that miR-137 and miR-29b bind to the 3'UTR region of GPx7 and inhibit the expression of this enzyme at the mRNA and protein level in all the human cells tested. However, no negative correlation between miR-137 nor miR-29b level and GPx7 was observed during adipogenesis. Despite the confirmed inhibition of GPx7 expression by miR-137 and miR-29b, the action of these two molecules in adipogenesis and mature adipocytes must be accompanied by other regulators.
谷胱甘肽过氧化物酶 7（GPx7）作为细胞内应激传感器/递质，在脂肪细胞分化和预防肥胖相关病理中发挥重要作用。因此，寻找控制 GPx7 表达的调节机制非常重要。由于 microRNAs（miRNAs）可能参与 GPx7 表达的调控，我们研究了与肥胖和脂肪生成有关的四种选定 miRNAs 对 GPx7 表达的抑制作用。通过转染选定的 miRNA 模拟物，在三种细胞模型（HEK293、SW480、AT-MSC）中测试了对 GPx7 表达的影响。通过荧光素酶基因报告基因测定法，跟踪了选定的 miRNAs 与 GPx7 的 3'UTR 的相互作用。此外，比较了来自四位人类供体的脂肪组织间充质干细胞（AT-MSC）和成熟脂肪细胞中 GPx7 和选定 miRNAs 的水平，并分析了在脂肪生成过程中这些水平的变化。我们的研究结果首次表明，miR-137 和 miR-29b 与 GPx7 的 3'UTR 区域结合，并在所有测试的人类细胞中抑制该酶的 mRNA 和蛋白水平表达。然而，在脂肪生成过程中，miR-137 或 miR-29b 的水平与 GPx7 之间没有观察到负相关。尽管 miR-137 和 miR-29b 证实抑制了 GPx7 的表达，但这两种分子在脂肪生成和成熟脂肪细胞中的作用必须伴随着其他调节剂。
