Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, College of Animal Science and Technology, Ningxia University, Yinchuan 750021, China.
Cells. 2023 Apr 17;12(8):1175. doi: 10.3390/cells12081175.
Circular RNAs (CircRNAs) are covalently closed-loop non-coding RNA (ncRNA) molecules present in eukaryotes. Numerous studies have demonstrated that circRNAs are important regulators of bovine fat deposition, but their precise mechanisms remain unclear. Previous transcriptome sequencing studies have indicated that circADAMTS16, a circRNA derived from the a disintegrin-like metalloproteinases with the thrombospondin motif 16 () gene, is high expressed in bovine adipose tissue. This gives a hint that the circRNA may be involved in the process of bovine lipid metabolism. In this study, the targeting relationship between circADAMTS16 and miR-10167-3p was verified using a dual-luciferase reporter assay. Then, the functions of circADAMTS16 and miR-10167-3p in bovine adipocytes were explored through gain-of-function and lose-of-function. The mRNA expression levels of genes were detected by real-time quantitative PCR (qPCR), and lipid droplet formation was phenotypically evaluated by Oil Red O staining. Cell proliferation and apoptosis were detected using CCK-8, EdU, and flow cytometry. We demonstrated that circADAMTS16 targeted binding to miR-10167-3p. The up-regulation of circADAMTS16 inhibited the differentiation of bovine preadipocytes, and the overexpression of miR-10167-3p promoted the differentiation of bovine preadipocytes. Meanwhile, CCK-8 and EdU results indicated that circADAMTS16 promoted adipocyte proliferation. Subsequently, flow cytometry analysis showed that circADAMTS16 promoted cell transition from G0/G1 phase to S phase, and inhibited cell apoptosis. However, up-regulation of miR-10167-3p inhibited cell proliferation and promoted apoptosis. Briefly, circADAMTS16 inhibited the differentiation and promotes the proliferation of bovine adipocytes by targeting miR-10167-3p during bovine fat deposition, which provides new insights into the mechanism of circRNAs regulation of beef quality.
环状 RNA（circRNAs）是真核生物中具有共价闭合环的非编码 RNA（ncRNA）分子。大量研究表明，circRNAs 是牛脂肪沉积的重要调节因子，但它们的确切机制尚不清楚。先前的转录组测序研究表明，环状 RNA circADAMTS16 是来自解整合素样金属蛋白酶与血栓反应蛋白 16 基因的一种 circRNA，在牛脂肪组织中高表达。这提示 circRNA 可能参与牛脂质代谢过程。在本研究中，通过双荧光素酶报告基因实验验证了 circADAMTS16 与 miR-10167-3p 的靶向关系。然后，通过功能获得和功能丧失实验探索了 circADAMTS16 和 miR-10167-3p 在牛脂肪细胞中的功能。通过实时定量 PCR（qPCR）检测基因的 mRNA 表达水平，并用油红 O 染色对脂滴形成进行表型评估。通过 CCK-8、EdU 和流式细胞术检测细胞增殖和凋亡。结果表明，circADAMTS16 靶向结合 miR-10167-3p。circADAMTS16 的上调抑制了牛前脂肪细胞的分化，而过表达 miR-10167-3p 促进了牛前脂肪细胞的分化。同时，CCK-8 和 EdU 结果表明 circADAMTS16 促进脂肪细胞增殖。随后，流式细胞术分析表明 circADAMTS16 促进细胞从 G0/G1 期向 S 期过渡，抑制细胞凋亡。然而，miR-10167-3p 的上调抑制了细胞增殖并促进了凋亡。综上所述，circADAMTS16 通过靶向 miR-10167-3p 抑制牛脂肪细胞分化并促进其增殖，从而促进牛脂肪沉积，为 circRNAs 调节牛肉质的机制提供了新的见解。
