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circRNA Profiling Reveals an Abundant circFUT10 that Promotes Adipocyte Proliferation and Inhibits Adipocyte Differentiation via Sponging let-7.
Molecular Therapy - Nucleic Acids ( IF 6.5 ) Pub Date : 2020-03-30 , DOI: 10.1016/j.omtn.2020.03.011
Rui Jiang 1 , Hui Li 2 , Jiameng Yang 1 , Xuemei Shen 1 , Chengchuang Song 1 , Zhaoxin Yang 1 , Xiaogang Wang 1 , Yongzhen Huang 1 , Xianyong Lan 1 , Chuzhao Lei 1 , Hong Chen 1
Affiliation  

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.



中文翻译:

circRNA分析揭示了丰富的circFUT10,可通过海绵体let-7促进脂肪细胞增殖并抑制脂肪细胞分化。

脂肪的发育受一系列复杂过程的调节,包括环状RNA(circRNA)在内的非编码RNA(ncRNA)在调节脂肪细胞的增殖和分化中起着重要作用。在这项研究中,我们分析了小牛和成年发育阶段牛脂肪组织中circRNA的表达,并检测到14,274个circRNA候选物。一些circRNA在两个发育阶段之间差异表达。我们表征了circFUT10,以其宿主基因FUT10(一种高表达和丰富的circRNA)命名。萤光素酶筛选,RNA结合蛋白免疫沉淀(RIP)分析,定量实时PCR和Western印迹分析表明circFUT10直接结合let-7c / let-e和PPARGC1B(过氧化物酶体增殖物激活受体γ共激活物1-β)被鉴定为let-7c的靶标。流式细胞仪,EdU(5-乙炔基-2'-脱氧尿苷)掺入,CCK-8(细胞计数试剂盒8)测定,油红O染色和Western印迹测定表明,circFUT10通过海绵化促进脂肪细胞增殖并抑制细胞分化let-7c。结果表明,circFUT10松懈7c的结合通过靶向促进细胞增殖并抑制细胞分化PPARGC1B牛脂肪细胞。

更新日期:2020-03-30
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