Posttranscriptional Interaction Between miR‐450a‐5p and miR‐28‐5p and STAT1 mRNA Triggers Osteoblastic Differentiation of Human Mesenchymal Stem Cells

Abstract

We demonstrate that the interaction between miR-450a-5p and miR-28-5p and signal transducer and activator of transcription 1 (STAT1) mRNA correlates with the osteoblastic differentiation of mesenchymal stem cells from human exfoliated deciduous teeth (shed cells). STAT1 negatively regulates runx-related transcription factor 2 (RUNX2), which is an essential transcription factor in this process. However, the elements that trigger osteoblastic differentiation and therefore pause the inhibitory effect of STAT1 need investigation. Usually, STAT1 can be posttranscriptionally regulated by miRNAs. To test this, we used an in vitro model system in which shed cells were chemically induced toward osteoblastic differentiation and temporally analyzed, comparing undifferentiated cells with their counterparts in the early (2 days) or late (7 or 21 days) periods of induction. The definition of the entire functional genome expression signature demonstrated that the transcriptional activity of a large set of mRNAs and miRNAs changes during this process. Interestingly, STAT1 and RUNX2 mRNAs feature contrasting expression levels during the course of differentiation. While undifferentiated or early differentiating cells express high levels of STAT1 mRNA, which was gradually downregulated, RUNX2 mRNA was upregulated toward differentiation. The reconstruction of miRNA-mRNA interaction networks allowed the identification of six miRNAs (miR-17-3p, miR-28-5p, miR-29b, miR-29c-5p, miR-145-3p, and miR-450a-5p), and we predicted their respective targets, from which we focused on miR-450a-5p and miR-28-5p STAT1 mRNA interactions, whose intracellular occurrence was validated through the luciferase assay. Transfections of undifferentiated shed cells with miR-450a-5p or miR-28-5p mimics or with miR-450a-5p or miR-28-5p antagonists demonstrated that these miRNAs might play a role as posttranscriptional controllers of STAT1 mRNA during osteoblastic differentiation. J. Cell. Biochem. 118: 4045-4062, 2017. © 2017 Wiley Periodicals, Inc.

MiR-596 inhibits osteoblastic differentiation and cell proliferation by targeting Smad3 in steroid-induced osteonecrosis of femoral head

“…It has been confirmed that the abnormal expression of miRNAs is closely related to stem cell differentiation [37]. For instance, miR-34a, miR-450a-5p, and miR-28- 5p have been revealed to be involved in regulating the osteoblastic differentiation of mesenchymal stem cells [38,39]. In BMSCs, the adipogenic and osteogenic differentiation can also be regulated by miR-383 [40], miR-20a-5p [41], and miR-20a-5p [41].…”

LncRNA‐PCAT1 targeting miR‐145‐5p promotes TLR4‐associated osteogenic differentiation of adipose‐derived stem cells

“…MiR‐145‐5p expression was decreased during osteogenic differentiation, and transfection of miR‐145 mimics in pluripotent mesenchymal precursor cells reduced the expression of osteogenic differentiation markers . The undifferentiated interaction networks of mesenchymal stem cells exhibited an increased miR‐145‐3p identification . In human osteoblast‐like MG‐63 cells, suppression of miR‐145 could stimulate the expression of osteoprotegerin, a soluble glycoprotein that reduced bone resorption and enhances bone formation .…”

“…16 The undifferentiated interaction networks of mesenchymal stem cells exhibited an increased miR-145-3p identification. 29 In human osteoblast-like MG-63 cells, suppression of miR-145 could stimulate the expression of osteoprotegerin, a soluble glycoprotein that reduced bone resorption and enhances bone formation. 30 MiR-145 also negatively affected the osteoblastic differentiation by targeting Cbfb, the transcription factors that are essential for bone formation.…”

MicroRNA‐29b regulatesDNAmethylation by targeting Dnmt3a/3b and Tet1/2/3 in porcine early embryo development

“…MicroRNAs (miRNAs) are 18–25 nucleotide noncoding RNAs that suppress the translation and stability of their target mRNAs generally through imperfect base‐pairing binding to the 3′ untranslated regions (3′ UTRs) of these target mRNAs (Rana, ). Research has increasingly suggested that diverse microRNAs (miRNAs) participate in numerous biological processes such as embryo development, stem cell pluripotency, differentiation, cell proliferation and apoptosis (Chen et al., , Dernowsek et al., ; Liu, Zhang, Sun, Su, & You, ; Wang et al., ; Zhang, Wang et al. ), but the functioning patterns and target genes of most miRNAs are still unknown.…”