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环状RNA分析揭示了一种丰富的环状FUT10,它通过海绵吸附let-7促进脂肪细胞增殖并抑制脂肪细胞分化。

circRNA Profiling Reveals an Abundant circFUT10 that Promotes Adipocyte Proliferation and Inhibits Adipocyte Differentiation via Sponging let-7.

作者信息

Jiang Rui, Li Hui, Yang Jiameng, Shen Xuemei, Song Chengchuang, Yang Zhaoxin, Wang Xiaogang, Huang Yongzhen, Lan Xianyong, Lei Chuzhao, Chen Hong

机构信息

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

Shaanxi Key Laboratory of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China; State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Animal Science and Technology, Guangxi University, Nanning 530004, China.

出版信息

Mol Ther Nucleic Acids. 2020 Jun 5;20:491-501. doi: 10.1016/j.omtn.2020.03.011. Epub 2020 Mar 30.

Abstract

Adipose development is regulated by a series of complex processes, and non-coding RNAs (ncRNAs), including circular RNAs (circRNAs), play important roles in regulating proliferation and differentiation of adipocytes. In this study, we profiled circRNA expression in cattle fat tissue during calf and adult developmental stages and detected 14,274 circRNA candidates. Some circRNAs are differentially expressed between two developmental stages. We characterized circFUT10, named for its host gene FUT10, a highly expressed and abundant circRNA. Luciferase screening, an RNA-binding protein immunoprecipitation (RIP) assay, quantitative real-time PCR, and western blotting assays indicated that circFUT10 directly binds let-7c/let-e, and PPARGC1B (peroxisome proliferator-activated receptor γ coactivator 1-β) is identified as a target of let-7c. Flow cytometry, EdU (5-ethynyl-2'-deoxyuridine) incorporation, a CCK-8 (cell counting kit-8) assay, oil red O staining, and western blotting assays demonstrated that circFUT10 promotes adipocyte proliferation and inhibits cell differentiation by sponging let-7c. The results demonstrate that circFUT10 binding of let-7c promotes cell proliferation and inhibits cell differentiation by targeting PPARGC1B in cattle adipocytes.

摘要

脂肪发育受一系列复杂过程调控,非编码RNA(ncRNA),包括环状RNA(circRNA),在调节脂肪细胞增殖和分化中发挥重要作用。在本研究中,我们分析了犊牛和成年牛发育阶段脂肪组织中circRNA的表达情况,共检测到14274个circRNA候选物。部分circRNA在两个发育阶段存在差异表达。我们鉴定了circFUT10,它以其宿主基因FUT10命名,是一种高表达且丰富的circRNA。荧光素酶筛选、RNA结合蛋白免疫沉淀(RIP)试验、定量实时PCR和蛋白质印迹试验表明,circFUT10直接结合let - 7c/let - e,且过氧化物酶体增殖物激活受体γ共激活因子1 - β(PPARGC1B)被鉴定为let - 7c的靶标。流式细胞术、EdU(5 - 乙炔基 - 2'-脱氧尿苷)掺入试验、CCK - 8(细胞计数试剂盒 - 8)试验、油红O染色和蛋白质印迹试验表明,circFUT10通过吸附let - 7c促进脂肪细胞增殖并抑制细胞分化。结果表明,在牛脂肪细胞中,circFUT10与let - 7c结合通过靶向PPARGC1B促进细胞增殖并抑制细胞分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb80/7163053/3be6865fbf7a/fx1.jpg

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