Key Laboratory of Hormones and Development (Ministry of Health)Tianjin Key Laboratory of Metabolic Diseases, Tianjin Metabolic Diseases Hospital & Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin, China
J Mol Endocrinol. 2018 Apr;60(3):225-237. doi: 10.1530/JME-17-0183. Epub 2018 Jan 18.
miR-20a-5p has recently been identified to induce adipogenesis of established adipogenic cell lines in our previous study. However, its role and molecular mechanisms in the regulation of adipocyte lineage commitment of bone marrow-derived stromal cells (BMSCs) still need to be explored. In this report, we demonstrated the expression of miR-20a-5p was promoted gradually during adipogenic differentiation in BMSCs. We also confirmed that miR-20a-5p has a positive function in the adipogenic differentiation of BMSCs by gain-of-function study with overexpression lentivirus or synthetic mimics of miR-20a-5p, and loss-of-function study with sponge lentivirus or synthetic inhibitor of miR-20a-5p. Dual luciferase reporter assay, GFP repression assay and Western blotting suggested Kruppel-like factor 3 () was a direct target of miR-20a-5p. Furthermore, siRNA-mediated silencing of recapitulated the potentiation of adipogenesis induced by miR-20a-5p overexpression, whereas enhanced expression of attenuated the effect of miR-20a-5p. As was reported to play an inhibitory role in adipogenesis at the initial stage of differentiation, the findings we present here indicate that miR-20a-5p promotes adipocyte differentiation from BMSCs by targeting and negatively regulating in the early phase during the procedure of adipogenesis.
miR-20a-5p 最近被鉴定为在我们之前的研究中诱导已建立的脂肪生成细胞系的脂肪生成。然而,其在骨髓基质细胞(BMSCs)脂肪细胞谱系分化中的作用和分子机制仍有待探索。在本报告中,我们证明了 miR-20a-5p 在 BMSCs 的脂肪生成分化过程中逐渐表达。我们还通过过表达慢病毒或 miR-20a-5p 的合成模拟物的功能获得研究,以及通过海绵慢病毒或 miR-20a-5p 的合成抑制剂的功能丧失研究,证实了 miR-20a-5p 在 BMSCs 的脂肪生成分化中具有正向作用。双荧光素酶报告基因检测、GFP 抑制检测和 Western blot 表明 Kruppel 样因子 3()是 miR-20a-5p 的直接靶标。此外,siRNA 介导的沉默 可重现 miR-20a-5p 过表达诱导的脂肪生成增强作用,而 表达增强则减弱了 miR-20a-5p 的作用。由于 被报道在分化的初始阶段在脂肪生成中起抑制作用,因此我们在这里的研究结果表明,miR-20a-5p 通过靶向和负调控脂肪生成过程早期的 来促进 BMSCs 的脂肪细胞分化。