Long Non-coding RNA H19 Regulates Porcine Satellite Cell Differentiation Through miR-140-5p/SOX4 and DBN1.
Long Non-coding RNA H19 Regulates Porcine Satellite Cell Differentiation Through miR-140-5p/SOX4 and DBN1.
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DOI：
10.3389/fcell.2020.518724
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发表时间：
2020
影响因子：
5.5
通讯作者：
Li C
中科院分区：
文献类型：
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作者：
Li J;Su T;Zou C;Luo W;Shi G;Chen L;Fang C;Li C
H19基因促进小鼠骨骼肌分化，但在猪中，H19基因调控肌肉生成的调控模型和机制在很大程度上是未知的。因此，需要确定H19在猪骨骼肌卫星细胞(PSCs)分化中的调控模式。我们观察到，H19基因沉默可以降低PSCs中肌生成素(MYOG)基因的表达，降低成肌分化(MYOD)和肌球蛋白重链(MYHC)的表达。因此，我们构建并测序了两个分化时间点H19基因敲除后PSCs的12个cDNA文库，以分析其转录组的差异。共鉴定出11,419个差异表达基因(Deg)。其中，通过生物信息学分析和蛋白质相互作用实验发现，SRY-box转录因子4(Sox4)和Drebrin 1(DBN1)是H19调控PSC分化的关键基因。功能分析表明，Sox4和DBN1促进PSC分化。从机制上讲，H19通过两条不同的途径调控PSC的分化。一方面，H19作为miR-140-5P的分子海绵，抑制PSCs的分化，从而调节Sox4的去阻抑作用。另一方面，H19通过直接与DBN1结合来调控PSC的分化。此外，MYOD与H19和DBN1的启动子结合。MYOD基因的敲除抑制了H19和DBN1的表达。我们确定了H19的功能，并提供了一个分子模型来阐明H19‘S在调节PSC分化中的作用。
The H19 gene promotes skeletal muscle differentiation in mice, but the regulatory models and mechanisms of myogenesis regulated by H19 are largely unknown in pigs. Therefore, the regulatory modes of H19 in the differentiation of porcine skeletal muscle satellite cells (PSCs) need to be determined. We observed that H19 gene silencing could decrease the expressions of the myogenin (MYOG) gene, myogenic differentiation (MYOD), and myosin heavy chain (MYHC) in PSCs. Therefore, we constructed and sequenced 12 cDNA libraries of PSCs after knockdown of H19 at two differentiation time points to analyze the transcriptome differences. A total of 11,419 differentially expressed genes (DEGs) were identified. Among these DEGs, we found through bioinformatics analysis and protein interaction experiment that SRY-box transcription factor 4 (SOX4) and Drebrin 1 (DBN1) were the key genes in H19-regulated PSC differentiation. Functional analysis shows that SOX4 and DBN1 promote PSC differentiation. Mechanistically, H19 regulates PSC differentiation through two different pathways. On the one hand, H19 functions as a molecular sponge of miR-140-5p, which inhibits the differentiation of PSCs, thereby modulating the derepression of SOX4. On the other hand, H19 regulates PSC differentiation through directly binding with DBN1. Furthermore, MYOD binds to the promoters of H19 and DBN1. The knockdown of MYOD inhibits the expression of H19 and DBN1. We determined the function of H19 and provided a molecular model to elucidate H19’s role in regulating PSC differentiation.