MiR-690, a Runx2-targeted miRNA, regulates osteogenic differentiation of C2C12 myogenic progenitor cells by targeting NF-kappaB p65

Abstract

BackgroundThe runt-related transcription factor 2 (Runx2) is a cell-fate-determining factor that controls osteoblast differentiation and bone formation. It has been previously demonstrated that microRNAs (miRNAs) play important roles in osteogenesis. However, the Runx2-regulated miRNAs that have been reported thus far are limited. In this study, we pursued to identify these miRNAs in Tet-on stable C2C12 cell line (C2C12/Runx2Dox subline).ResultsMicroarray analysis revealed that alterations in miRNA expression occur with 54 miRNAs. Among these miRNAs, miR-690 was identified as a positive regulator of Runx2-induced osteogenic differentiation of C2C12 cells through gain- and loss-of-function assays. Expression of miR-690 is induced by Runx2, which binds directly to the putative promoter of mir-690 (Mirn690). The miR-690 proceeds to inhibit translation of the messenger RNA encoding the nuclear factor kappa B (NF-κB) subunit p65 whose overexpression inhibits Runx2-induced osteogenic differentiation of C2C12 cells. Interleukin-6 (IL-6), a downstream target of NF-κB pathway, is upregulated by p65 overexpression but significantly downregulated during this differentiation process. Furthermore, overexpression of IL-6 impedes the expression of osteocalcin, a defined marker of late osteoblast differentiation.ConclusionsTogether, our results suggest that the miR-690 transactivated by Runx2 acts as a positive regulator of Runx2-induced osteogenic differentiation by inactivating the NF-κB pathway via the downregulation of the subunit p65.Electronic supplementary materialThe online version of this article (doi:10.1186/s13578-016-0073-y) contains supplementary material, which is available to authorized users.

The microRNA-10a/ID3/RUNX2 axis modulates the development of Ossification of Posterior Longitudinal Ligament

“…Recently, various miRNAs that regulate osteogenic differentiation have been identified. For example, miR-690 activated by RUNX2 positively regulates RUNX2-induced osteogenic differentiation by inactivating the NF-κB pathway via the downregulation of subunit p65 7 , whereas TNF-α-induced NF-kB activation upregulates microRNA-150-3p and inhibits osteogenesis of mesenchymal stem cells by targeting β-catenin 8 . These results suggest that osteogenesis is tightly controlled by miRNAs, and miRNA regulation may be an upstream mediator of the osteogenic process.…”

MicroRNA‐3064‐3p regulates the differentiation of cementoblasts through targeting DKK1

“…In particular, numerous studies have published that miRNAs play critical roles in osteogenic differentiation . For instance, miR‐690 was found to regulate osteogenic differentiation of C2C12 cells through targeting nuclear factor‐kappaB p65 . Besides, miR‐144‐3p regulates C3H10T1/2 cells through modulating Smad4, miR‐194 modulates osteoblast differentiation by targeting signal transducer and activator of transcription 1‐mediated runt‐related gene 2 nuclear translocation, and miR‐214 has a crucial role in inhibiting bone formation by directly targeting activating transcription factor 4 .…”

Pseudomonas Aeruginosa Secreted Protein PA3611 Promotes Bronchial Epithelial Cells Epithelial-Mesenchymal Transition Through TGF-β1 Inducing p38/miRNA/NF-κB Pathway

“…It is well known that NF-κB is an important intersection for a variety of signaling pathways, and many factors play roles in NF-κB activation, in recent years, the activating effect of microRNA (miRNA) has attracted much attention [42,43]. miRNA is a kind of single-stranded small molecule RNA with a length of about 21~25 nucleotides that widely exists in eukaryotes.…”