Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Sciences and Technologies, Northwest A&F University, Yangling 712100, China.
Acta Biochim Biophys Sin (Shanghai). 2020 Dec 11;52(11):1227-1235. doi: 10.1093/abbs/gmaa111.
miRNAs are a small class of noncoding RNAs that perform biological functions by regulating the stability or translation of target genes in various biological processes. This study illustrated the role of miR-10a-5p, which is relatively enriched in adipose tissues, using primary mouse preadipocytes as model. With elevated miR-10a-5p expression, the proliferative ability of mouse preadipocytes was significantly enhanced, indicated by increased EdU+ cells and G1/S transition, accompanied by upregulated Cyclin B, Cyclin D and PCNA and downregulated p21 and p27. Meanwhile, the adipogenic differentiation was significantly attenuated by elevated miR-10a-5p, supported by Oil Red O staining and suppressed PPARγ and aP2 expression. Furthermore, Map2k6 and Fasn were predicted to be the target genes of miR-10a-5p in silico, and dual luciferase reporter assay confirmed the direct targeting effects. Western blot analysis results showed that miR-10a-5p specially reduced Map2k6 expression at the proliferative stage without affecting Fasn expression, while significantly restrained Fasn expression with unchanged Map2k6 expression during adipogenic differentiation. Taken together, these results revealed a potential role of miR-10a-5p in adipogenesis and in the treatment of obesity.
miRNAs 是一类小的非编码 RNA,通过调节靶基因的稳定性或翻译在各种生物过程中发挥生物学功能。本研究以原代小鼠前体脂肪细胞为模型,说明了相对富含脂肪组织的 miR-10a-5p 的作用。随着 miR-10a-5p 表达的升高,小鼠前体脂肪细胞的增殖能力显著增强,表现在 EdU+细胞和 G1/S 期转换增加,同时细胞周期蛋白 B、D 和 PCNA 上调,p21 和 p27 下调。同时,miR-10a-5p 的升高显著抑制了脂肪生成分化,表现在油红 O 染色减少和 PPARγ 和 aP2 表达受抑制。此外,Map2k6 和 Fasn 被预测为 miR-10a-5p 的靶基因,双荧光素酶报告基因实验证实了其直接靶向作用。Western blot 分析结果表明,miR-10a-5p 在增殖期特异性降低 Map2k6 表达,而不影响 Fasn 表达,而在脂肪生成分化过程中,miR-10a-5p 显著抑制 Fasn 表达,而不改变 Map2k6 表达。综上所述,这些结果揭示了 miR-10a-5p 在脂肪生成和肥胖治疗中的潜在作用。