MiRNA‐1271‐5p regulates osteogenic differentiation of human bone marrow‐derived mesenchymal stem cells by targeting forkhead box O1 (FOXO1)

Abstract

Forkhead box O1 (FOXO1) is a key regulator of osteogenesis. The aim of this study was to identify the mechanisms of microRNAs (miRNAs) targeting FOXO1 in osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). Three miRNA target prediction programs were used to search for potential miRNAs that target FOXO1. Quantitative real‐time polymerase chain reaction was conducted to detect the expression of miR‐1271‐5p and FOXO1 during osteogenic differentiation. Target gene prediction and screening, luciferase reporter assay was used to verify the downstream target gene of miR‐1271‐5p. The expression levels of FOXO1 and Runx2 were detected by RT‐qPCR and Western blot analysis. Alkaline phosphatase (ALP) activity and matrix mineralization were detected by biochemical methods. The expression levels of Runx2, ALP, and osteocalcin were detected by RT‐qPCR. Our results showed that miR‐1271‐5p was downregulated during osteogenic induction. And the expression levels of miR‐1271‐5p were higher in osteoporotic tissues than that in adjacent nonosteoporotic tissues. The expression levels of FOXO1 were lower in osteoporotic tissues than that in adjacent nonosteoporotic tissues. And a negative correlation was found between miR‐1271‐5p and FOXO1 in osteoporotic tissues. Overexpression of miR‐1271‐5p downregulated FOXO1 and inhibited osteogenic differentiation in hMSCs. Overexpression of miR‐1271‐5p downregulated the expression of osteogenic markers and reduced ALP activity. In addition, ectopic expression of FOXO1 reversed the effect of miR‐1271‐5p on osteogenic differentiation. In conclusion, miR‐1271‐5p functioned as a therapeutic target of osteogenic differentiation in hMSCs by inhibiting FOXO1, which provides valuable insights into the use of miR‐1271‐5p as a target in the treatment of osteoporosis and other bone metabolic diseases.

Identification of Potential Osteoporosis miRNA Biomarkers Using Bioinformatics Approaches

“…The result showed that 13 miRNAs were remarkably highly expressed in PBMC of mice with osteoporosis. Among the 13 miRNAs, the expression trends of miR-1271-5p, miR-135a-5p, miR-135b-5p, miR-153-3p, miR-223-3p, miR-27a-3p, miR-370-3p, and miR-9-5p in the current study were in line with their trends in various studies [ 5 , 7 , 41 – 46 ]. Our result revealed that the expression of the 13 miRNAs was positively correlated with the progression of osteoporosis.…”

“…As noncoding small RNAs, miRNAs can posttranscriptionally modulate the expression of specific genes, thus affecting various kinds of biological processes [4]. A number of studies on miRNAs affecting osteoporosis exhibited that miRNAs can regulate various genes and inhibit proliferation and differentiation of osteoblasts, thereby causing osteoporosis [5][6][7]. Meanwhile, works of literature also reported that predictive models for fracture risk due to osteoporosis can be constructed via miRNAs [8].…”

Molecular Mechanism of Long Noncoding RNA SNHG14 in Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells through the NEDD4L/FOXA2/PCP4 Axis

“…11 Stem Cells International NEDD4L as an E3 ubiquitin-protein ligase mediates ubiquitination modification to regulate the protein level 54], and our bioinformatics data suggested FOXA2 was a target of NEDD4L. FOX family members are essential for bone metabolism and function as regulators of OD of MSCs [28,[55][56][57]. FOXA2 is identified as a critical transcription factor involved in the process of MSCs in the repair of the injured pancreas, intestinal and hepatic tissues, and neurons [58][59][60][61].…”

Time series clustering analysis of genes during osteogenic differentiation of human mesenchymal stem cells

“…Various studies have indicated the importance of TWIST1 (Miraoui et al, 2010;Quarto et al, 2015;Fan et al, 2022) and LEP (Xu et al, 2016;Zhao et al, 2016) in the regulation of osteogenic differentiation: LEP reportedly alters the differentiation fate of chondrogenic progenitor cells and triggers their senescence (Zhao et al, 2016), and promotes osteoblast differentiation of BMSCs (Xu et al, 2016); meanwhile, inhibiting the expression of TWIST1 could promote the osteogenic differentiation of periodontal MSCs (Xu et al, 2019) and adipose tissue-derived stem cells (Quarto et al, 2015). FOXO1 transcription factor, a member of the Fox family, is mainly involved in cell proliferation, differentiation, angiogenesis and other biological processes, and multiple studies have also reported the involvement of FOXO1 during osteogenic differentiation (Hao et al, 2018;Chen et al, 2019;Yang et al, 2021). In our study, these four genes were all markedly up-regulated in osteogenic induction samples in comparison with uninduced hMSC samples, suggesting that they play vital roles in osteogenic differentiation of hMSCs, as reported in the above studies.…”