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高表达的 miR-10a-5p 分别通过靶向 Map2k6 和 Fasn 促进细胞周期和抑制成脂分化。

Elevated miR-10a-5p facilitates cell cycle and restrains adipogenic differentiation via targeting Map2k6 and Fasn, respectively.

机构信息

Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Sciences and Technologies, Northwest A&F University, Yangling 712100, China.

出版信息

Acta Biochim Biophys Sin (Shanghai). 2020 Dec 11;52(11):1227-1235. doi: 10.1093/abbs/gmaa111.

Abstract

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.

摘要

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。

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