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bta-miR-204 在脂肪细胞增殖、分化和凋亡中的调控作用。

Role of bta-miR-204 in the regulation of adipocyte proliferation, differentiation, and apoptosis.

机构信息

Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, People's Republic of China.

Department of Animal Science, College of Agriculture and Natural Resources, University of Wyoming, Laramie, Wyoming.

出版信息

J Cell Physiol. 2019 Jul;234(7):11037-11046. doi: 10.1002/jcp.27928. Epub 2019 Jan 30.

Abstract

Adipocyte growth and development are complex and precisely orchestrated processes. Several microRNAs have been identified as critical regulators of the adipocyte growth and development. Recently, bta-miR-204 was found to be involved in adipogenesis; however, the underlying molecular mechanism involved in bta-miR-204-mediated regulation of proliferation, differentiation, and apoptosis of adipocytes is not fully understood or elucidated. In this study, quantitative real-time polymerase chain reaction (qRT-PCR), Cell Counting Kit-8, EdU, flow cytometer, Oil Red O staining, and the western blot assays were used to assess the role of bta-miR-204 in adipocyte growth and development. Overexpression of bta-miR-204 had no significant effect on 3T3-L1 cell proliferation. The forced expression of bta-miR-204 promoted 3T3-L1 cell differentiation. Meanwhile, overexpression of bta-miR-204 upregulated the expression of Bax and downregulated the expression of Bcl-2 both at messenger RNA and protein levels, which suggested that bta-miR-204 can promote 3T3-L1 cell apoptosis. Using bioinformatic analysis, dual-luciferase reporter system and qRT-PCR, TGFBR2, and ELOVL6 were identified as the direct target genes of bta-miR-204. Therefore, our study provides a novel insight into the role of bta-miR-204 in the regulation of adipocyte growth and development, which may provide a novel therapeutic alternative against obesity.

摘要

脂肪细胞的生长和发育是一个复杂而精确的过程。有几种 microRNA 被鉴定为脂肪细胞生长和发育的关键调节因子。最近,bta-miR-204 被发现参与脂肪生成;然而,bta-miR-204 介导的脂肪细胞增殖、分化和凋亡调节的潜在分子机制尚未完全阐明。在这项研究中,使用定量实时聚合酶链反应(qRT-PCR)、细胞计数试剂盒-8、EdU、流式细胞仪、油红 O 染色和 Western blot 分析来评估 bta-miR-204 在脂肪细胞生长和发育中的作用。bta-miR-204 的过表达对 3T3-L1 细胞的增殖没有显著影响。bta-miR-204 的强制表达促进了 3T3-L1 细胞的分化。同时,bta-miR-204 的过表达上调了 Bax 的表达,下调了 Bcl-2 的表达,mRNA 和蛋白质水平均如此,这表明 bta-miR-204 可以促进 3T3-L1 细胞凋亡。通过生物信息学分析、双荧光素酶报告系统和 qRT-PCR,鉴定出 TGFBR2 和 ELOVL6 是 bta-miR-204 的直接靶基因。因此,我们的研究为 bta-miR-204 在调节脂肪细胞生长和发育中的作用提供了新的见解,这可能为肥胖症的治疗提供新的选择。

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