miR-10167-3p 以 TCF7L1 为靶标,抑制牛脂肪细胞分化,促进牛脂肪细胞增殖。

IF 3.4 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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引用次数: 0

摘要

微RNA(miRNA)广泛参与各种致脂过程,包括脂肪细胞的增殖和分化、脂滴的形成以及脂肪细胞特异性基因的激活。本研究旨在利用 RNA-seq 技术研究 miR-10167-3p 高表达下牛前脂肪细胞的基因表达谱,并验证其下游靶基因对牛前脂肪细胞增殖和分化的功能。首先,RNA-seq鉴定了573个差异表达基因(DEGs),其中243个下调,330个上调。然后,京都基因和基因组百科全书(KEGG)分析显示,15.19%的DEGs富集在与脂质代谢相关的通路中。同时,双荧光素酶报告基因检测验证了 miR-10167-3p 与 TCF7L1 的靶结合关系。在 TCF7L1 高表达和 RNA 干扰的脂肪细胞中进行了多项实验,以评估 TCF7L1 的功能。利用 qPCR 和 western 印迹分别检测了增殖、分化和凋亡标记基因的 mRNA 和蛋白表达;利用油红 O、尼罗河红和波迪比染色检测了脂滴的合成;利用 EdU 检测了脂肪细胞的增殖;利用流式细胞术检测了细胞凋亡。结果显示,TCF7L1过表达会抑制牛前脂肪细胞的分化和凋亡,促进其增殖,而对其进行RNA干扰则会得到相反的结果。这些结果可为后续研究牛脂肪沉积的分子机制提供参考。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
miR-10167-3p targets TCF7L1 to inhibit bovine adipocyte differentiation and promote bovine adipocyte proliferation

MicroRNAs (miRNAs) are widely involved in various lipogenic processes, including adipocyte proliferation and differentiation, lipid droplet formation, and adipocyte-specific gene activation. The present study aimed to investigate the gene expression profiles of bovine preadipocytes under high miR-10167-3p expression using the RNA-seq technique and to verify the functions of its downstream target genes on the proliferation and differentiation of bovine preadipocytes. First, RNA-seq identified 573 differentially expressed genes (DEGs), of which 243 were downregulated and 330 were upregulated. Then, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that 15.19% of the DEGs were enriched in pathways related to lipid metabolism. Meanwhile, dual-luciferase reporter gene assay verified the target-binding relationship between miR-10167-3p and TCF7L1. The function of TCF7L1 was assessed using several experiments in adipocytes with high TCF7L1 expression and RNA interference. The mRNA and protein expression of proliferation, differentiation, and apoptosis marker genes were detected using qPCR and western blot, respectively; lipid droplet synthesis was detected using oil red O, Nile red, and bodipy staining; adipocyte proliferation was detected by EdU; and apoptosis was detected using flow cytometry. The results revealed that TCF7L1 overexpression inhibited bovine preadipocyte differentiation and apoptosis and promoted their proliferation, with opposite results obtained with its RNA interference. These results may provide a reference for the subsequent investigation of the molecular mechanism of bovine fat deposition.

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来源期刊
Genomics
Genomics 生物-生物工程与应用微生物
CiteScore
9.60
自引率
2.30%
发文量
260
审稿时长
60 days
期刊介绍: Genomics is a forum for describing the development of genome-scale technologies and their application to all areas of biological investigation. As a journal that has evolved with the field that carries its name, Genomics focuses on the development and application of cutting-edge methods, addressing fundamental questions with potential interest to a wide audience. Our aim is to publish the highest quality research and to provide authors with rapid, fair and accurate review and publication of manuscripts falling within our scope.
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