miR-133a-3p inhibits the osteogenic differentiation of bone marrow mesenchymal stem cells by regulating ankyrin repeat domain 44

Abstract

In this study, we aimed to identify the specific microRNAs (miRNAs) that are involved in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) from ovariectomized (OVX) mice, and to further explore the mechanism by which these miRNAs regulate osteogenic differentiation. Based on the existing studies, the expression of seven miRNAs in BMSCs from OVX mice was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of miR-133a-3p and osteogenesis-related genes (runt-related transcription factor 2 (Runx2), Osterix, alkaline phosphatase (ALP), and osteopontin) in BMSCs treated with miR-133a-3p mimics or inhibitors was detected by qRT-PCR or Western blotting. Osteogenesis efficiency was determined using ALP and alizarin red staining. The effector-target relationship between miR-133a-3p and ankyrin repeat domain 44 (ANKRD44) was confirmed by bioinformatics and a dual luciferase assay. Among the seven selected miRNAs, miR-133a-3p expression was significantly increased in BMSCs from OVX mice. Overexpression of miR-133a-3p dramatically inhibited the expression of osteogenesis-related genes in BMSCs and reduced ALP activity and mineralization. However, these processes were markedly ameliorated upon miR-133a-3p inhibition. Moreover, miR-133a-3p appeared to target ANKRD44, and the ANKRD44 expression was negatively regulated by miR-133a-3p. Furthermore, ANKRD44 upregulation eliminated the anti-osteogenic differentiation effects of miR-133a-3p in BMSCs. Thus, our results indicated that miR-133a-3p inhibits the osteogenic differentiation of BMSCs by suppressing ANKRD44.

Signature of circulating small non-coding RNAs during early fracture healing in mice

“…In callus tissue, miR-133a-3p levels correlated negatively with Met expression ( Bourgery et al, 2021 ). Recent data indicates that overexpression of miR-133a-3p inhibits osteogenesis of bone marrow mesenchymal stem cells by regulating ankyrin repeat domain 44 ( Ankrd44 ) ( Li et al, 2021 ). Based on these previous studies, we propose that miR-133a-3p functions as a regulator of bone remodelling via targeting P13K/AKT to regulate osteogenic differentiation.…”

MicroRNA expression in JAG1/Notch-activated periodontal ligament stem cells

“…It is noteworthy that both upregulation as well as downregulation of several miRNAs and differential patterns of osteogenesis have been reported in different Jagged1-exposed cell types by a number of researchers [ 35 – 37 ]. Thus, Li et al [ 35 ] found overexpression of mhesiR-133a-3p markedly inhibited the expression of osteogenesis-associated genes such as RUNX2 and OSX , reduced ALP activity and mineralization by targeting ankyrin repeat domain 44 (ANKRD44) in BMMSCs. Similarly, Deng et al [ 36 ] observed significant inhibition of osteogenic differentiation, cell migration, and cell proliferation in MC3T3-E1 cells due to miR-210-3p.…”

SP1 regulates BMSC osteogenic differentiation through the miR-133a-3p/MAPK3 axis

“…Furthermore, miRNAs can modulate the progression of various diseases, including OP [ 39 ]. Mao et al found that miR-133a-3p regulates osteogenic differentiation by regulating the repeat domain of ankyrin [ 40 ]. MiR-133a-3p is also targeted by the lncRNA MEG3 to inhibit osteogenic differentiation during the progression of postmenopausal OP [ 29 ].…”