Downregulation of long non-coding RNA H19 promotes P19CL6 cells proliferation and inhibits apoptosis during late-stage cardiac differentiation via miR-19b-modulated Sox6

Abstract

BackgroundRegulating cardiac differentiation to maintain normal heart development and function is very important. At present, biological functions of H19 in cardiac differentiation is not completely clear.MethodsTo explore the functional effect of H19 during cardiac differentiation. Expression levels of early cardiac-specific markers Nkx-2.5 and GATA4, cardiac contractile protein genes α-MHC and MLC-2v were determined by qRT-PCR and western lot. The levels of lncRNA H19 and miR-19b were detected by qRT-PCR. We further predicted the binding sequence of H19 and miR-19b by online softwares starBase v2.0 and TargetScan. The biological functions of H19 and Sox6 were evaluated by CCK-8 kit, cell cycle and apoptosis assay and caspase-3 activity.ResultsThe expression levels of α-MHC, MLC-2v and H19 were upregulated, and miR-19b was downregulated significantly in mouse P19CL6 cells at the late stage of cardiac differentiation. Biological function analysis showed that knockdown of H19 promoted cell proliferation and inhibits cell apoptosis. H19 suppressed miR-19b expression and miR-19b targeted Sox6, which inhibited cell proliferation and promoted apoptosis in P19CL6 cells during late-stage cardiac differentiation. Importantly, Sox6 overexpression could reverse the positive effects of H19 knockdown on P19CL6 cells.ConclusionDownregulation of H19 promoted cell proliferation and inhibited cell apoptosis during late-stage cardiac differentiation by regulating the negative role of miR-19b in Sox6 expression, which suggested that the manipulation of H19 expression could serve as a potential strategy for heart disease.

The novel regulatory role of lncRNA‐miRNA‐mRNA axis in cardiovascular diseases

“…During the late stage of cardiac differentiation of P19CL6 cells, miR‐19b was negatively regulated by H19, as demonstrated by luciferase activity assay and qRT‐PCR. The overexpression of H19 significantly inhibited cell proliferation and promoted cell apoptosis via modulating miR‐19b and its target sox6, while the down‐regulation of H19 reversed these effects …”

“…It has been gradually demonstrated that H19 may modulate a variety of cell biological functions by sequestering different miRNAs; H19 can serve as not only as a sponge of miRNAs, such as let‐7, miR‐103/107 and miR‐19b, in cell differentiation, necrosis and proliferation, but also as a precursor of miRNAs in the regulation of cardiomyocyte hypertrophy. The miR‐675 was validated to be located in the first exon of H19, and both H19 and miR‐675 were up‐regulated in hypertrophic heart tissue.…”

“…The overexpression of H19 significantly inhibited cell proliferation and promoted cell apoptosis via modulating miR-19b and its target sox6, while the down-regulation of H19 reversed these effects. 32 Cardiomyocyte autophagy is an evolutionarily conserved process in face of stress stimuli, but abnormal autophagy contributes to cell death. 33 The lncRNA AK079427, named APF, played a crucial role in the regulation of cardiomyocyte autophagy by modulating miR-188-3p and its target ATG7.…”

PTENP1 inhibits the growth of esophageal squamous cell carcinoma by regulating SOCS6 expression and correlates with disease prognosis

“…It also provides miR‐Function and ceRNA‐Function web tools to predict the function of ncRNAs (miRNAs, lncRNAs, pseduogenes) and protein‐coding genes from the miRNA‐mediated (ceRNA) regulatory networks . StarBase was commonly used in studying lncRNA‐miRNA‐mRNA network …”

The Cytomegalovirus Enhancer Induces an Immediate Response to the Myosin Light Chain 2v Promoter during P19CL6 Cell Differentiation

“…Similar to these cardiac MHC isoforms, cardiac myosin light chain 2 (MLC-2v) is also transcribed during the course of differentiation [7]. When the 250 base pair (bp) promoter for the rat cardiac MLC-2v gene was connected upstream of the gene for green fluorescent protein (GFP) together with the enhancer portion of the cytomegalovirus (CMV) immediate early promoter, and the resulting reporter plasmid was stably introduced into P19CL6 cells, expression of GFP was limited to developing cardiac myocytes [8].…”