Circular RNA circMYL1 inhibit proliferation and promote differentiation of myoblasts by sponging miR-2400

Abstract

Background Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs (ncRNAs) involved in regulating skeletal muscle development by sponging microRNAs (miRNAs). This study aimed to define the circMYL1 molecular mechanisms in the regulation of bovine myogenesis and to disclose its regulatory mechanism through miR-2400 interaction. Methods The potential role of circMYL1 was identified in the proliferation of bovine myoblast through mRNA and protein expression of proliferation marker genes (PCNA, CyclinD1, and CDK2), cell counting kit-8 (CCK8) assay, flow cytometry analysis, and EdU assay. Analysis of the expression of differentiation marker genes (MyoD, MyoG, and MYH2) and immunofluorescence of Myosin heavy chain (MyHC) was used to assess cell differentiation. Results The proliferation analysis revealed that circMYL1 inhibited the proliferation of bovine primary myoblast. Furthermore, the differentiation analysis demonstrated that circMYL1 promoted the differentiation of bovine primary myoblast. The luciferase screening and RNA immunoprecipitation (RIP) assays found that circMYL1 could have interaction with miR-2400. Additionally, we demonstrated that miR-2400 promoted proliferation and inhibited differentiation of bovine primary myoblast, while circMYL1 may eliminate the effects of miR-2400, as showed by rescue experiments. Conclusions Our results revealed that a novel circular RNA of circMYL1 could inhibit proliferation and promote differentiation of myoblast by sponging miR-2400.

Circular RNA circUSP13 sponges miR‐29c to promote differentiation and inhibit apoptosis of goat myoblasts by targeting IGF1

“…The FISH assay was performed according to a previously described procedure, 25 with minor modifications. Cy3‐labeled probes against 18S and circUSP13 were designed and synthesized by Genepharma (Shanghai, China).…”

Circular RNA regulation of fat deposition and muscle development in cattle

“…Therefore, circRILPL1 is positively correlated with muscle proliferation and differentiation in vitro and can inhibit cell apoptosis, thus promoting the growth of bovine myoblasts. CircMYL1 interacts with miR‐2400 to inhibit myoblast proliferation and promote differentiation through the competitive adsorption of miR‐2400, which promotes cell proliferation by targeting MYOG (Elnour et al., 2021). MYOG , a member of the myogenic regulatory factors family, induces terminal differentiation of skeletal muscle (Liu et al., 2020a,b).…”