MiR-27a-3p promotes the osteogenic differentiation by activating CRY2/ERK1/2 axis

Abstract

Background Osteoporosis seriously disturbs the life of people. Meanwhile, inhibition or weakening of osteogenic differentiation is one of the important factors in the pathogenesis of osteoporosis. It was reported that miR-27a-3p reduced the symptoms of osteoporosis. However, the mechanism by which miR-27a-3p in osteogenic differentiation remains largely unknown. Methods To induce the osteogenic differentiation in MC3T3-E1 cells, cells were treated with osteogenic induction medium (OIM). RT-qPCR was used to evaluate the mRNA expression of miR-27a-3p and CRY2 in cells. The protein levels of CRY2, Runt-related transcription factor 2 (Runx2), osteopontin (OPN), osteocalcin (OCN) and the phosphorylation level of extracellular regulated protein kinases (ERK) 1/2 in MC3T3-E1 cells were evaluated by western blotting. Meanwhile, calcium nodules and ALP activity were tested by alizarin red staining and ALP kit, respectively. Luciferase reporter gene assay was used to analyze the correlation between CRY2 and miR-27a-3p. Results The expression of miR-27a-3p and the phosphorylation level of ERK1/2 were increased by OIM in MC3T3-E1 cells, while CRY2 expression was decreased. In addition, OIM-induced increase of calcified nodules, ALP content and osteogenesis-related protein expression was significantly reversed by downregulation of miR-27a-3p and overexpression of CRY2. In addition, miR-27a-3p directly targeted CRY2 and negatively regulated CRY2. Meanwhile, the inhibitory effect of miR-27a-3p inhibitor on osteogenic differentiation was reversed by knockdown of CRY2 or using honokiol (ERK1/2 signal activator). Furthermore, miR-27a-3p significantly inhibited the apoptosis of MC3T3-E1 cells treated by OIM. Taken together, miR-27a-3p/CRY2/ERK axis plays an important role in osteoblast differentiation. Conclusions MiR-27a-3p promoted osteoblast differentiation via mediation of CRY2/ERK1/2 axis. Thereby, miR-27a-3p might serve as a new target for the treatment of osteoporosis.

BMSC-derived exosomal miR-27a-3p and miR-196b-5p regulate bone remodeling in ovariectomized rats

“…In accordance with the present results, previous studies have demonstrated that miR-27a-3p enhanced osteogenic differentiation of MC3T3-E1 preosteoblasts ( Ren et al, 2021 ; Xu et al, 2018a ; Xu et al, 2018b ). However, several studies have shown that miR-27a-3p inhibits the osteogenic differentiation of MC3T3-E1 preosteoblasts or BMSCs ( Xu et al, 2020 ; Zeng et al, 2021 ; Furesi et al, 2022 ; Zhang et al, 2017 ).…”

MicroRNA sequence and function analysis in peri‐implantitis and periodontitis: An animal study

“…For instance, miR‐27a‐3p was downregulated in both PI vs. PD and PI vs. PIC. MiR‐27a‐3p can directly target CRY2, promoting the phosphorylation of ERK1/2, inhibiting apoptosis, and promoting the osteogenic differentiation of MC3T3‐E1 cells 25 . Our previous research found that miR‐27a‐3p was downregulated in dogs with peri‐implantitis compared with healthy dogs, 17 and miR‐27a‐3p may target DKK2 and SFRP1 to promote reosseointegration in the regenerative treatment of peri‐implantitis 14 .…”

“…MiR-27a-3p can directly target CRY2, promoting the phosphorylation of ERK1/2, inhibiting apoptosis, and promoting the osteogenic differentiation of MC3T3-E1 cells. 25 Our previous research found that miR-27a-3p was downregulated in dogs with peri-implantitis compared with healthy dogs, 17 and miR-27a-3p may target DKK2 and SFRP1 to promote reosseointegration in the regenerative treatment of peri-implantitis. 14 Recent studies suggested that miR-27a-3p promotes autophagy and apoptosis in IL-1β-treated articular chondrocytes in osteoarthritis through PI3K/AKT/mTOR signaling.…”

Assessment of microRNA profiles in small extracellular vesicles isolated from bovine colostrum with different immunoglobulin G concentrations

“…No differentially abundant miRNA was identified based on differential expression analysis except for miR-27a-3p, which was more abundant in high-compared with low-IgG samples (P = 0.028; Figure 2C). This miR-27a-3p showed to promote osteogenic differentiation in pre-osteoblast (MC3T3-E1) cells (Ren et al, 2021), suggesting a role in the osteogenesis of neonatal calves.…”