MicroRNA-409-3p promotes osteoblastic differentiation via activation of Wnt/β-catenin signaling pathway by targeting SCAI

Abstract

Osteogenic differentiation is an important process of new bone formation, microRNA-409-3p (miR-409-3p) has been reported to be up-regulated in the osteogenic differentiation of human bone marrow mesenchymal stem cells (MSCs). The present study aimed to investigate the regulatory effect of miR-409-3p on osteogenic differentiation of MSCs and its molecular mechanism. The expression of miR-409-3p in osteoblast (human skull osteoblast, HCO) and bone marrow-derived MSCs (MSC-A, MSC-B, MSC-U) were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The binding of miR-409-3p to suppressor of cancer cell invasion (SCAI) in MSC-B was investigated by performing a dual-luciferase reporter gene assay. MSC-B was selected to transfect with miR-409-3p analog/complementary sequence (cs), miR-409-3p analog + SCAI and miR-409-3p cs + small interfering (si)-SCAI, as well as control, respectively. The alkaline phosphatase (ALP) activity, Alizarin Red staining, and the expression of osteogenic markers (ALP, osteocalcin (OCN), osteopontin (OPN), runt-related transcription factor 2 (RUNX2)) in MSC-B during osteoblastic differentiation were tested by RT-qPCR and Western blotting, respectively. Additionally, the Wnt/β-catenin pathway was inhibited by dickkopf-related protein 1 (DKK-1) to get the roles of miR-409-3p during the osteoblastic differentiation of MSC-B when transfected with miR-409-3p analog. The expression of miR-409-3p in HCO was higher than that in these three MSCs and showed an increasing time-dependent trend on the 0 and 21st day of osteoblastic differentiation. MiR-409-3p directly regulated SCAI by targeting SCAI 3′UTR. Further, miR-409-3p suppressed SCAI expression, but SCAI up-regulation suppressed the osteoblastic differentiation, as well as reduced the relative mRNA/protein expression of Wnt/β-catenin signaling pathway-related genes (Axis inhibition protein 1 (AXIN1), β-catenin, Lymphoid Enhancer Binding Factor 1, Cellular-myelocytomatosis (c-myc) and cyclin D1). Importantly, disruption of Wnt signaling also blocked miR-409-3p induced osteoblastic differentiation of MSCs. Therefore, miR-409-3p promotes osteoblastic differentiation through the activation of the Wnt/β-catenin pathway by down-regulating SCAI expression.

The effects of Omarigliptin on promoting osteoblastic differentiation

“…We further measured the expression levels of important factors involved in the regulation of osteoblastic differentiation, such as BMP-2 [ 20 ], Ocn [ 21 ], and ALP [ 22 ]. We found that BMP-2, Ocn, and ALP ( Figure 3a-b ) were significantly upregulated in osteogenic medium-cultured MC3T3‑E1 cells at both the mRNA and protein levels, and pronouncedly further upregulated by 20 μM Omarigliptin, revealing the regulatory function of Omarigliptin on osteoblastic differentiation is associated with these factors.…”

miRNAs Related to Different Processes of Fracture Healing: An Integrative Overview

“…MiR-409 has shown to enhance osteogenic differentiation by activating the Wnt-signalling pathway. MiR-409 directly targeted Suppressor of Cancer Cell Invasion, thereby reducing its expression, whereas Suppressor of Cancer Cell Invasion upregulation showed to decrease osteoblastic differentiation and reduce protein expression of Wnt-signalling related genes, such as β-catenin, c-myc, and cyclin D1 (107). A glucose regulated miRNA, miR-451, enhanced osteoblastogenesis in vitro and in vivo, by suppressing Odd Skipped Related 1 whilst enhancing RUNX2 and BMP-4 expression (67).…”

miR-409-3p Regulated by GATA2 Promotes Cardiac Fibrosis through Targeting Gpd1

“…miR-409-3p, an unexplored candidate, has been primarily investigated in oncology. For instance, miR-409-3p promotes epithelial to mesenchymal transition and tumor growth in prostate cancer and also accelerates osteoblastic differentiation through the Wnt/ β -catenin pathway [ 17 , 18 ], a crucial pathway in regulating cardiac fibrosis. Previous clinical study discovered its potential value in distinguishing atrial fibrillation [ 19 ].…”