Lnc-GD2H Promotes Proliferation by Forming a Feedback Loop With c-Myc and Enhances Differentiation Through Interacting With NACA to Upregulate Myog in C2C12 Myoblasts

Abstract

In the present study, the roles of a novel long non-coding RNA (lncRNA), lnc-GD2H, in promoting C2C12 myoblast proliferation and differentiation and muscle regeneration were investigated by quantitative polymerase chain reaction, western blotting, Cell Counting Kit-8, 5-ethynyl-2′-deoxyuridine (EdU), immunofluorescence staining, luciferase reporter, mass spectrometry, pulldown, chromatin immunoprecipitation, RNA immunoprecipitation assay, wound healing assays, and cardiotoxin (CTX)-induced muscle injury assays. It was observed that lnc-GD2H promoted myoblast proliferation as evidenced by the enhancement of the proliferation markers c-Myc, CDK2, CDK4, and CDK6, percentage of EdU-positive cells, and rate of cell survival during C2C12 myoblast proliferation. Additional experiments confirmed that c-Myc bound to the lnc-GD2H promoter and regulated its transcription. lnc-GD2H promoted cell differentiation with enhanced MyHC immunostaining as well as increased expression of the myogenic marker genes myogenin (Myog), Mef2a, and Mef2c during myoblast differentiation. Additional assays indicated that lnc-GD2H interacted with NACA which plays a role of transcriptional regulation in myoblast differentiation, and the enrichment of NACA at the Myog promoter was impaired by lnc-GD2H. Furthermore, inhibition of lnc-GD2H impaired muscle regeneration after CTX-induced injury in mice. lnc-GD2H facilitated the expression of proliferating marker genes and formed a feedback loop with c-Myc during myoblast proliferation. In differentiating myoblasts, lnc-GD2H interacted with NACA to relieve the inhibitory effect of NACA on Myog, facilitating Myog expression to promote differentiation. The results provide evidence for the role of lncRNAs in muscle regeneration and are useful for developing novel therapeutic targets for muscle disorders.

RRM2 promotes the proliferation of chicken myoblasts, inhibits their differentiation and muscle regeneration

“…Previous studies have shown that MyoG and MyoD are both myogenic regulatory factors (MRFs), which play important roles in myoblast differentiation and fusion and can be used as mouse C2C12 myoblast differentiation marker genes ( Zhang et al, 2018 ; Chen et al, 2021b ). In chickens and mice, MyHC is highly expressed during myogenic differentiation and myotube formation and is also a differentiation marker gene in myogenesis ( Chen et al, 2019b ; Xu et al, 2019 ; Sakai et al, 2022 ).…”

lncRNA GPRC5D-AS1 as a ceRNA inhibits skeletal muscle aging by regulating miR-520d-5p

“…Among these factors, MyoG plays a crucial role in the growth, development and regeneration of skeletal muscle [50,51]. Additionally, Mef2c is a member of the myocyte enhancer factor 2 (Mef2) family, which involves in regulating the expression of muscle regulatory factors and skeletal muscle-specific transcription of these factors [52].…”

Transcriptome Analysis of mRNA and lncRNA Related to Muscle Growth and Development in Gannan Yak and Jeryak

“…lncRNA, which is characterized by a length exceeding a certain value, has emerged as crucial a regulator of gene function and expression levels in this context [25][26][27][28]. According to recent research, lncRNA plays a crucial role in the regulation of skeletal muscle growth and development [29][30][31]. In the present study, a total of 304 DELs and 1819 DEMs were identified, among which 132 lncRNAs and 1081 mRNAs were found to be down-regulated, while 172 lncRNAs and 738 mRNAs were up-regulated.…”