Long Non-coding RNA H19 Regulates Porcine Satellite Cell Differentiation Through miR-140-5p/SOX4 and DBN1

Abstract

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.

LncRNA H19 governs mitophagy and restores mitochondrial respiration in the heart through Pink1/Parkin signaling during obesity

“…Results from studies using different cell types support that H19 is mainly, even exclusively, expressed in the cytoplasm 43 – 45 . Based on this subcellular location, competing endogenous RNA regulation is the most reported mechanism of action of H19, followed by direct binding to transcription regulators or other proteins 46 , 47 .…”

Roles of the H19/microRNA‑675 axis in the proliferation and epithelial‑mesenchymal transition of human cutaneous squamous cell carcinoma cells

“…An increasing number of studies have reported an association between lncRNAs and cSCC ( 27 ). Accumulating evidence has revealed that H19, as a cancer-related lncRNA, served an important role in physiological activities and pathological mechanisms ( 16 , 28 , 29 ). For example, H19 was reported to act as an oncogene in bladder, gastric, breast and thyroid cancer, in addition to glioma ( 15 , 30 – 33 ).…”

LncRNA TCONS_00323213 Promotes Myogenic Differentiation by Interacting with PKNOX2 to Upregulate MyoG in Porcine Satellite Cells

“…H19 , a well-known imprinted lncRNA [ 9 ], is expressed in animal skeletal muscle and heart tissues and exerts important functions [ 10 ]. H19 can inhibit insulin-like growth factor 2 ( IGF2) transcription by binding to polycomb repressive complex 2 (PRC2) [ 11 ] and recruit TAR DNA-binding protein 43 (TDP43) and drebrin 1 (DBN1) to induce differentiation of PSCs [ 12 , 13 ]. The lncRNA metastasis-associated lung adenocarcinoma transcript 1 ( Malat1) functions via the suppressor of variation 39 homolog 1 (Suv39h1) to repress the expression of the target gene myogenic differentiation factor ( MyoD) [ 14 ].…”