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. 2021 Dec 1;21(4):568-576.

miR-21 regulates osteogenic and adipogenic differentiation of BMSCs by targeting PTEN

Yongtao Zhou  1 , Hongwang Qiao  1 , Lili Liu  2 , Ping Dong  3 , Fangxu Zhu  4 , Jiawen Zhang  5 , Liping Liu  1 , Li Liu  1
Affiliations

miR-21 regulates osteogenic and adipogenic differentiation of BMSCs by targeting PTEN

Yongtao Zhou et al. J Musculoskelet Neuronal Interact. .

Abstract

Objective: To explore the effects and mechanism of miR-21 on the osteogenic/adipogenic differentiation of mouse BMSCs.

Methods: The bilateral ovaries of C57BL/6J mice (n=24) were removed to construct an osteoporosis model. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-21, osteogenic/adipogenic genes, and PTEN. ALP and ARS and ORO staining were used to detect the formation of calcium nodules and lipid droplets in BMSCs. Western blot was used to detect the expression of PTEN.

Results: miR-21 was significantly down-regulated in osteoporotic mice. The expression of miR-21 was significantly up-regulated after the osteogenic induction of BMSCs, and the expression of miR-21 was significantly down-regulated after the adipogenic induction. Overexpression of miR-21 significantly promoted the osteogenic differentiation of BMSCs and inhibits the adipogenic differentiation of BMSCs.

Conclusion: MiR-21 can promote osteogenic differentiation of BMSCs and inhibit their adipogenic differentiation by negatively regulating PTEN.

Keywords: Adipose differentiation; BMSCs; Osteogenic Differentiation; Osteoporosis; miRNA.

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Conflict of interest statement

The authors have no conflict of interest.

Figures

Figure 1
Figure 1
Expression of miR-21 in OP mice. (A-D) Detection of bone-related indexes by microCT, SMI = structure model index; (E) qRT-PCR detection of the expression of miR-21 in bone tissue of OP mice; (F) qRT-PCR detection of the expression of miR-21 in mouse BMSCs in OP group.
Figure 2
Figure 2
miR-21 expression changes before and after differentiation in BMSCs of mice. (A) qRT-PCR detection of the expression of miR-21 before and after osteogenesis of mouse BMSCs; (B) qRT-PCR detection of the expression of miR-21 before and after adipogenesis of mouse BMSCs.
Figure 3
Figure 3
Effects of overexpression of miR-21 on osteogenic differentiation of mouse BMSCs. (A) Observation of miR-21 expression in mouse BMSCs by qRT-PCR; (B) Detection of calcium nodule formation in BMSCs by alkaline phosphatase staining; (C) Manifestation of calcium nodule emergence in BMSCs by Alizarin red staining; (D) Detection of the expression degrees of osteogenesis-related genes Runx2, ALP, OCN and OPN in mouse BMSCs by qRT-PCR.
Figure 4
Figure 4
Effect of overexpression of miR-21 on adipogenic differentiation of mouse BMSCs. (A) Detection of the formation of lipid droplets in BMSCs by oil red O staining; (B) Quantitative results of oil red O staining; (C) Detection of the expression levels of adipogenesis related genes CEBPα, CEBPβ, FABP4 and PPARγ in mouse BMSCs by qRT-PCR.
Figure 5
Figure 5
miR-21 negatively regulates PTEN expression. (A) The binding site of miR-21 and PTEN; (B) Luciferase reporter assay was used to detect the regulatory association between miR-21 and PTEN; (C) Detection of PTEN mRNA expression by qRT-PCR; (D) Detection of the protein expression of PTEN was observed by western blot; (E) Statistical results of western blot.
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