Retinoid acid-induced microRNA-31-5p suppresses myogenic proliferation and differentiation by targeting CamkIIδ

Abstract

BackgroundWe previously reported that Wnt5a/CaMKIIδ (calcium/calmodulin-dependent protein kinase II delta) pathway was involved in the embryonic tongue deformity induced by excess retinoic acid (RA). Our latest study found that the expression of miR-31-5p, which was predicted to target the 3′UTR of CamkIIδ, was raised in the RA-treated embryonic tongue. Thus, we hypothesized that the excess RA regulated Wnt5a/CaMKIIδ pathway through miR-31-5p in embryonic tongue.MethodsC2C12 myoblast line was employed as an in vitro model to examine the suppression of miR-31-5p on CamkIIδ expression, through which RA impaired the myoblast proliferation and differentiation in embryonic tongue.ResultsRA stimulated the expression of miR-31-5p in both embryonic tongue and C2C12 myoblasts. Luciferase reporter assay confirmed that the 3′UTR of CamkIIδ was a target of miR-31-5p. MiR-31-5p mimics disrupted CamkIIδ expression, C2C12 proliferation and differentiation as excess RA did, while miR-31-5p inhibitor partially rescued these defects in the presence of RA.ConclusionsExcess RA can stimulate miR-31-5p expression to suppress CamkIIδ, which represses the proliferation and differentiation of tongue myoblasts.Electronic supplementary materialThe online version of this article (doi:10.1186/s13395-017-0126-x) contains supplementary material, which is available to authorized users.

miR-27b-3p Was Involved in Retinoic Acid-induced Abnormal Early Myogenic Differentiation of C2C12 Cells via Targeting CaMKIIδ

“…3A) and DTNA 15) for the seed sequence of miR-27b-3p. We also found another miRNA, miR-31-5p could also complementary match with the 3' UTR of CamkIIδ 14) , but the effects of both miR-27b-3p targeted to DTNA and miR-31-5p to CaMKIIδ were mainly displayed in the late stages of C2C12 myogenic diff erentiation 14,15) . So, whether miR-27b-3p targeted CaMKIIδ aff ected the other processes of myogenesis attracted our attention.…”

“…To explore the molecular mechanism for this, we subsequently did miRNA microarray and identified the dysregulated miRNAs in RA-induced abnormal tongue 15) and studies in this paper, we found that CaMKIIδ was a downstream target of miR-27b-3p. Furthermore, our latest studies also indicated that miR-27b-3p and miR-31-5p could target DTNA 15) and CaMKIIδ 14) respectively at late myogenic diff erentiation stages, implying that miR-27b-3p controls different targets and CaMKIIδ could be regulated by numbers of miRNAs at different stages during myogenic diff erentiation process. These were consistent with the characters and complex regulatory network of miRNAs.…”

“…In the meantime, the expression of CaMKIIδ was decreased at the same stages 14) . Bioinformation analysis using TargetScan, MiRanda, and MicroCosm predicted that a conserve complementary site in both the 3'UTRof CamkIIδ (Fig.…”

“…The procedure of RNA oligonucleotides transfection was carried out as described in our previous study 14) . The RNA oligonucleotides' sequences are listed in Table 1.…”

Effect of IC50 dose of retinol on metabolomics of RAW264.7 cells

“…A previous study demonstrated the involvement of the Wnt5a/CaMKII pathway in retinoic acid (RA)-induced developmental abnormalities in mouse genioglossus [17]. Retinoic acid (10 µM) inhibits the proliferation and differentiation of C2C12 cells by upregulating the expression of miR-31-5p and miR-27b-3p and downregulating the expression of CamkIIδ and DTNA [18,19]. Exogenous RA (3 µM) promotes the apoptosis and inhibits the proliferation of epithelial cells lining the labial cervical loop by downregulating the expression of the mesenchymal factor Fgf10, thereby negatively regulating the homeostasis of the incisor stem cell niche.…”