Zhang Wei Wei, Sun Xiao Feng, Tong Hui Li, Wang Ya Hui, Li Shu Feng, Yan Yun Qin, Li Guang Peng
The Laboratory of Cell and Development, Northeast Agricultural University, Mucai Street 59, Xiangfang District, Harbin, 150030 Heilongjiang China.
College of Life Sciences and Agriculture & Forestry, Qiqihar University, Qiqihar, Heilongjiang 161006 China.
Cell Mol Biol Lett. 2016 Jul 28;21:8. doi: 10.1186/s11658-016-0009-x. eCollection 2016.
The differentiation of skeletal muscle-derived satellite cells (MDSCs) is important in controlling muscle growth, improving livestock muscle quality, and healing of muscle-related disease. MicroRNAs (miRNAs) are a class of gene expression regulatory factors, which play critical roles in the regulation of muscle cell differentiation. This study aimed to compare the expression profile of miRNAs in MDSC differentiation, and to investigate the miRNAs which are involved in MDSC differentiation.
Total RNA was extracted from MDSCs at three different stages of differentiation (MDSC-P, MDSC-D1 and MDSC-D3, representing 0, 1 and 3 days after differentiation, respectively), and used to construct small RNA libraries for RNA sequencing (RNA-seq).
The results showed that in total 617 miRNAs, including 53 novel miRNA candidates, were identified. There were 9 up-expressed, 165 down-expressed, and 15 up-expressed, 145 down-expressed in MDSC-D1 and MDSC-D3, respectively, compared to those in MDSC-P. Also, 17 up-expressed, 55 down-expressed miRNAs were observed in MDSC-D3 compared to those in MDSC-D1. All known miRNAs belong to 237 miRNA gene families. Furthermore, we observed some sequence variants and base edits of the miRNAs. GO and KEGG pathway analysis showed that the majority of target genes regulated by miRNAs were involved in cellular metabolism, pathways in cancer, actin cytoskeleton regulation and the MAPK signaling pathway. Regarding the 53 novel miRNAs, there were 7 up-expressed, 31 down-expressed, and 8 up-expressed, 26 down-expressed in MDSC-D1 and MDSC-D3, respectively, compared to those in MDSC-P. The expression levels of 12 selected miRNA genes detected by RT-qPCR were consistent with those generated by deep sequencing.
This study confirmed the authenticity of 564 known miRNAs and identified 53 novel miRNAs which were involved in MDSC differentiation. The identification of novel miRNAs has significantly expanded the repertoire of bovine miRNAs and could contribute to advances in understanding muscle development in cattle.
骨骼肌来源的卫星细胞（MDSCs）的分化在控制肌肉生长、改善家畜肌肉品质以及肌肉相关疾病的愈合中起着重要作用。微小RNA（miRNAs）是一类基因表达调控因子，在肌肉细胞分化的调控中发挥关键作用。本研究旨在比较miRNAs在MDSC分化过程中的表达谱，并研究参与MDSC分化的miRNAs。
从处于三个不同分化阶段的MDSCs（MDSC-P、MDSC-D1和MDSC-D3，分别代表分化后0、1和3天）中提取总RNA，并用于构建小RNA文库进行RNA测序（RNA-seq）。
结果显示，共鉴定出617个miRNAs，包括53个新的miRNA候选物。与MDSC-P相比，MDSC-D1和MDSC-D3中分别有9个上调、165个下调，以及15个上调、145个下调。此外，与MDSC-D1相比，MDSC-D3中观察到17个上调、55个下调的miRNAs。所有已知的miRNAs属于237个miRNA基因家族。此外，我们还观察到了miRNAs的一些序列变异和碱基编辑。GO和KEGG通路分析表明，miRNAs调控的大多数靶基因参与细胞代谢、癌症通路、肌动蛋白细胞骨架调控和MAPK信号通路。对于53个新的miRNAs，与MDSC-P相比，MDSC-D1和MDSC-D3中分别有7个上调、31个下调，以及8个上调、26个下调。通过RT-qPCR检测的12个选定miRNA基因的表达水平与深度测序结果一致。
本研究证实了564个已知miRNAs的真实性，并鉴定出53个参与MDSC分化的新miRNAs。新miRNAs的鉴定显著扩展了牛miRNAs的种类，有助于推动对牛肌肉发育的理解。
