Bta-miR-885 promotes proliferation and inhibits differentiation of myoblasts by targeting MyoD1.

The proliferation and differentiation of myoblasts are essential for the regeneration and development of skeletal muscles. However, the process of skeletal muscle development in cattle is complex and needs to be further investigated. The microRNAs (miRNAs) are endogenous, small noncoding RNAs that play a critical role during skeletal muscle development. In this study, we evaluated the function of miR-885 in muscle development in cattle. The results found that the expression of miR-885 was gradually upregulated during myoblast proliferation, whereas progressively downregulated during myoblast differentiation. The overexpression of miR-885 promoted cell proliferation of myoblast in cattle. Moreover, we further noted that the overexpression miR-885 triggered the expression level of various marker genes involved in cell proliferation, including proliferating cell nuclear antigen (PCNA), cyclin-dependent kinase 2 (CDK2), and cyclin B1 (CCNB1). Furthermore, it was observed that overexpression of miR-885 inhibited cell differentiation, and significantly decreased messenger RNA and protein expression levels of myogenic differentiation 1 (MyoD1) and myogenin (MyoG) in primary bovine myoblasts. Moreover, the miR-885 inhibitor revealed that miR-885 inhibited cell proliferation and promoted cell differentiation. In addition, the overexpression of miR-885 markedly decreased MyoD1 expression in primary bovine myoblasts. The luciferase reporter assay, quantitative real-time polymerase chain reaction, and western blot (WB) further indicated that miR-885 directly binding to 3' UTR of MyoD1 gene during transcriptional regulation. Conclusively, these results signified that miR-885 could be critical for the proliferation and differentiation in primary bovine myoblast cells by targeting the MyoD1 gene in cattle.