MicroRNA-27b-3p Targets the Myostatin Gene to Regulate Myoblast Proliferation and Is Involved in Myoblast Differentiation.

Genxi Zhang,Tao Zhang,Xinchao Zhang,Guojun Dai,Kaizhou Xie,Pengfei Wu,Mingliang He,Tingting Li,Kaizhi Zhou,Jinyu Wang

doi:10.3390/cells10020423

Abstract

microRNAs play an important role in the growth and development of chicken embryos, including the regulation of skeletal muscle genesis, myoblast proliferation, differentiation, and apoptosis. Our previous RNA-seq studies showed that microRNA-27b-3p (miR-27b-3p) might play an important role in regulating the proliferation and differentiation of chicken primary myoblasts (CPMs). However, the mechanism of miR-27b-3p regulating the proliferation and differentiation of CPMs is still unclear. In this study, the results showed that miR-27b-3p significantly promoted the proliferation of CPMs and inhibited the differentiation of CPMs. Then, myostatin (MSTN) was confirmed to be the target gene of miR-27b-3p by double luciferase reporter assay, RT-qPCR, and Western blot. By overexpressing and interfering with MSTN expression in CPMs, the results showed that overexpression of MSTN significantly inhibited the proliferation and differentiation of CPMs. In contrast, interference of MSTN expression had the opposite effect. This study showed that miR-27b-3p could promote the proliferation of CPMs by targeting MSTN. Interestingly, both miR-27b-3p and MSTN can inhibit the differentiation of CPMs. These results provide a theoretical basis for further understanding the function of miR-27b-3p in chicken and revealing its regulation mechanism on chicken muscle growth.

Highlights

In recent years, with more and more attention paid to microRNA, it has become the focus of biological research
The results showed that the expression level of miR-27b-3p in the process of differentiation was lower than that in the proliferation stage, which indicated that miR-27b-3p was involved in chicken primary myoblasts (CPMs) differentiation (Figure 3a)
The rescue experiments showed that the overexpression of miR-27b-3p and MSTN in CPMs could down-regulate the role of miR27b-3p in proliferation. These results indicate that MSTN is the target gene of miR-27b-3p, and miR-27b-3p regulates CPMs proliferation by targeting MSTN

Summary

Introduction

With more and more attention paid to microRNA (miRNA), it has become the focus of biological research. MiRNA is a class of endogenous noncoding singlechain small molecule RNA, about 21–26 nucleotides [2]. It cannot encode proteins, but it can participate in post-transcriptional regulation. MiRNA has a wide range of biological functions and participates in various cell processes, including proliferation, differentiation, apoptosis, metabolism, invasion, metastasis, and drug resistance [4,5]. In the process of skeletal muscle differentiation, the regulation of genes in the myomiRs family is essential. Many non-myomiRs family genes play a key role, which indicates that miRNAs play an important role in skeletal muscle growth and development [9,10]

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