环状 RNA 家族 10 通过海绵吸附 miR-133a 减少成肌细胞的增殖并促进其分化。
CircFUT10 reduces proliferation and facilitates differentiation of myoblasts by sponging miR-133a.
机构信息
Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, P.R. China.
Institute for Conservation and Utilization of Agro-Bioresources in Dabie Mountains, College of Life Sciences, Xinyang Normal University, Xinyang, Henan, P.R. China.
出版信息
J Cell Physiol. 2018 Jun;233(6):4643-4651. doi: 10.1002/jcp.26230. Epub 2018 Jan 15.
Circular RNAs (circRNAs) have been identified in various tissues and cell types from human, monkey, porcine, and mouse. However, knowledge on circRNAs in bovine muscle development is limited. We downloaded and analyzed the circRNAs sequencing data of bovine skeletal muscle tissue, and further characterized the role of a candidate circRNA (circFUT10) in muscle development. Quantitative real-time PCR (qPCR) and Western blot assays were used to confirm the expression of genes involved in myoblasts differentiation and proliferation. Flow cytometry was performed to assess cell cycle distribution and cell apoptosis. EdU incorporation and CCK-8 assay were performed to demonstrate cell proliferation. We demonstrated that circFUT10 was highly (but differentially) expressed in embryonic and adult skeletal muscle tissue. circFUT10 induced bovine primary myoblasts differentiation and increased the expression of MyoD, MyoG, and MyhC in mRNA and protein levels. circFUT10 increased the number of myoblasts in the G0/G1 phase of the cell cycle, and decreased the proportion of cells in the S-phase. circFUT10 inhibited the proliferation of myoblasts and promoted them apoptosis. Via a luciferase screening assay, circFUT10 is observed to sponge to miR-133a with three potential binding sites. Specifically, we show that circFUT10 regulated myoblasts differentiation and cell survival by directly binding to miR-133a and inhibiting miR-133a activity. Modulation of circFUT10 expression in muscle tissue may emerge as a potential target in breeding strategies attempting to control muscle development in cattle.
环状 RNA(circRNAs)已在人类、猴子、猪和鼠的各种组织和细胞类型中被鉴定出来。然而,关于牛肌肉发育中的 circRNAs 的知识有限。我们下载并分析了牛骨骼肌组织的 circRNAs 测序数据,并进一步表征了候选 circRNA(circFUT10)在肌肉发育中的作用。定量实时 PCR(qPCR)和 Western blot 分析用于确认参与成肌细胞分化和增殖的基因的表达。流式细胞术用于评估细胞周期分布和细胞凋亡。EdU 掺入和 CCK-8 测定用于证明细胞增殖。我们证明了 circFUT10 在胚胎和成年骨骼肌组织中高度(但差异)表达。circFUT10 诱导牛原代成肌细胞分化,并增加 MyoD、MyoG 和 MyhC 在 mRNA 和蛋白水平的表达。circFUT10 增加了细胞周期 G0/G1 期的成肌细胞数量,并降低了 S 期细胞的比例。circFUT10 抑制成肌细胞的增殖并促进其凋亡。通过荧光素酶筛选测定,观察到 circFUT10 与三个潜在结合位点结合到 miR-133a 上。具体来说,我们表明 circFUT10 通过直接与 miR-133a 结合并抑制 miR-133a 活性来调节成肌细胞分化和细胞存活。在试图控制牛肌肉发育的育种策略中,调节肌肉组织中的 circFUT10 表达可能成为一个潜在的目标。