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