title: Identification of miR-2400 gene as a novel regulator in skeletal muscle satellite cells proliferation by targeting MYOG gene.
abstract: MicroRNAs play critical roles in skeletal muscle development as well as in regulation of muscle cell proliferation and differentiation. Previous study in our laboratory showed that the expression level of miR-2400, a novel and unique miRNA from bovine, had significantly changed in skeletal muscle-derived satellite cells (MDSCs) during differentiation, however, the function and expression pattern for miR-2400 in MDSCs has not been fully understood. In this report, we firstly identified that the expression levels of miR-2400 were down-regulated during MDSCs differentiation by stem-loop RT-PCR. Over-expression and inhibition studies demonstrated that miR-2400 promoted MDSCs proliferation by EdU (5-ethynyl-2' deoxyuridine) incorporation assay and immunofluorescence staining of Proliferating cell nuclear antigen (PCNA). Luciferase reporter assays showed that miR-2400 directly targeted the 3' untranslated regions (UTRs) of myogenin (MYOG) mRNA. These data suggested that miR-2400 could promote MDSCs proliferation through targeting MYOG. Furthermore, we found that miR-2400, which was located within the eighth intron of the Wolf-Hirschhorn syndrome candidate 1-like 1 (WHSC1L1) gene, was down-regulated in MDSCs in a direct correlation with the WHSC1L1 transcript by Clustered regularly interspaced palindromic repeats interference (CRISPRi). In addition, these observations not only provided supporting evidence for the codependent expression of intronic miRNAs and their host genes in vitro, but also gave insight into the role of miR-2400 in MDSCs proliferation.
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China; College of Life Sciences and Agriculture & Forestry, Qiqihar University, Qiqihar, Heilongjiang 161006, China.
authors.forename: Wei Wei
authors.initials: WW
authors.lastname: Zhang
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
authors.forename: Hui Li
authors.initials: HL
authors.lastname: Tong
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
authors.forename: Xiao Feng
authors.initials: XF
authors.lastname: Sun
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Hu
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Yang
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China.
authors.forename: Shu Feng
authors.initials: SF
authors.lastname: Li
authors.affiliation: The Laboratory of Cell and Development, Northeast Agricultural University, Harbin, Heilongjiang 150030, China. Electronic address: yanyunqin@sohu.com.
authors.forename: Yun Qin
authors.initials: YQ
authors.lastname: Yan
authors.affiliation: The Key Laboratory of Mammal Reproductive Biology and Biotechnology Ministry of Education, Inner Mongolia University, Hohhot 010021, China.
authors.forename: Guang Peng
authors.initials: GP
authors.lastname: Li
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D002417:Cattle; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics*; D002478:Cells, Cultured; D017931:DNA Primers; D015536:Down-Regulation; D035683:MicroRNAs / Q000235:genetics*; D018008:Myogenin / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D017931:DNA Primers; D035683:MicroRNAs; D018008:Myogenin; D012333:RNA, Messenger
keywords: CRISPRi; Myogenin; Proliferation; Skeletal muscle-derived satellite cells; miR-2400