miR-460b-5p promotes proliferation and differentiation of chicken myoblasts and targets RBM19 gene
文献类型: 外文期刊
第一作者: Zhou, Kai-Zhi
作者: Zhou, Kai-Zhi;Wu, Peng-Fei;Ling, Xuan-Ze;Zhang, Jin;Wang, Qi-Fan;Zhang, Xin-Chao;Zhang, Tao;Zhang, Gen-Xi;Xue, Qian;Han, Wei
作者机构:
关键词: miRNA; skeletal muscle; growth and development; chicken
期刊名称:POULTRY SCIENCE ( 影响因子:4.4; 五年影响因子:4.4 )
ISSN: 0032-5791
年卷期: 2024 年 103 卷 1 期
页码:
收录情况: SCI
摘要: The meat production of broilers is crucial to economic benefits of broiler industries, while the slaughter performance of broilers is directly determined by skeletal muscle development. Hence, the broiler breed-ing for growth traits shows a great importance. As a kind of small noncoding RNA, microRNA (miRNA) can reg-ulate the expression of multiple genes and perform a wide range of regulation in organisms. Currently, more and more studies have confirmed that miRNAs are closely associated with skeletal muscle development of chickens. Based on our previous miR-seq analysis (accession num-ber: PRJNA668199), miR-460b-5p was screened as one of the key miRNAs probably involved in the growth regula-tion of chickens. However, the regulatory effect of miR-460b-5p on the development of chicken skeletal muscles is still unclear. Therefore, miR-460b-5p was further used for functional validation at the cellular level in this study. The expression pattern of miR-460b-5p was investigated in proliferation and differentiation stages of chicken pri-mary myoblasts. It was showed that the expression level of miR-460b-5p gradually decreased from the proliferation stage (GM 50%) to the lowest at 24 h of differentiation. As differentiation proceeded, miR-460b-5p expression increased significantly, reaching the highest and stabiliz-ing at 72 h and 96 h of differentiation. Through mRNA quantitative analysis of proliferation marker genes, CCK-8 and Edu assays, miR-460b-5p was found to significantly facilitate the transition of myoblasts from G1 to S phase and promote chicken myoblast proliferation. mRNA and protein quantitative analysis of differentiation marker genes, as well as the indirect immunofluorescence results of myotubes, revealed that miR-460b-5p significantly stimulated myotube development and promote chicken myoblast differentiation. In addition, the target relation-ship was validated for miR-460b-5p according to the dual-luciferase reporter assay and mRNA quantitative analysis, which indicates that miR-460b-5p was able to regulate RBM19 expression by specifically binding to the 30 UTR of RBM19. In summary, miR-460b-5p has posi-tive regulatory effects on the proliferation and differentia-tion of chicken myoblasts, and RBM19 is a target gene of miR-460b-5p.
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