The influence of MicroRNA‐150 in Osteoblast Matrix Mineralization

Abstract

This study investigated the influence of miR-150 expression on osteoblast matrix mineralization and its mechanisms. The mouse osteoblast cell line MC3T3-E1 was used as an in vitro model of bone formation. On the fifth day of mineralization, transfection experiments using agomiR-150, agomiR-NC, antagomiR-150 antagomiR-NC, and mock groups were set up to test the effects of miR-150 in MC3T3-E1 model. The mRNA and protein levels of OC, ALP, type I collagen, and OPN were measured by qRT-PCR and ELISA. Matrix mineralization was detected by alizarin red S (ARS) staining and flow cytometry was employed to quantify apoptosis in each group. RT-PCR and Western blot were applied to detect the expression of target gene MMP14. Our results demonstrated that the endogenous expression levels of miR-150, OC, ALP, type I collagen, and OPN in MC3T3-E1 cells increased steadily. Exogenous expressions of agomiR-150 and antagomiR-150 can significantly up-/down-regulate, respectively, the expression level of miR-150 in MC3T3-E1 cells. Compared with the mock group, higher expression levels of OC, ALP, type I collagen, and OPN mRNA were observed in the agomiR-150 group, while lower mRNA expression levels of OC, ALP, type I collagen, and OPN were found in the antagomiR-150 group. Based on these results, potential miR-150 targeted genes are discussed. Our results showed that miR-150 supports the osteoblastic phenotype related to osteoblast function and bone mineralization. Thus, miR-150 may have potential therapeutic applications in promoting bone formation in certain disease settings, such as in osteoporosis and in elderly patients.

Potential of delphinidin-3-rutinoside extracted from Solanum melongena L. as promoter of osteoblastic MC3T3-E1 function and antagonist of oxidative damage

“…Specific sets of primers for ALP, collagen type 1, alpha 1 (ColA1), osteocalcin (OC), OPG and RANKL. -catenin cDNAs were designed and synthesized (Sigma-Aldrich), according to Zhang et al [33] and Dong et al [34]. Quantitative PCR was performed in CFX96 Bio-Rad using 20 μl of total volume.…”

The Regulatory Roles of MicroRNAs in Bone Remodeling and Perspectives as Biomarkers in Osteoporosis

“…MiR-223 is reported to be an effective therapeutic target to relieve advanced glycation end products-induced damage to osteoblasts in diabetes mellitus [ 125 ]. MiR-150 could promote bone formation; the agomir of miR-150 may be considered as a novel therapy for osteoporosis [ 126 ]. MiR-214 is a key regulator of osteoclast differentiation, which may be considered as a promising therapeutic target for osteoporosis [ 106 ].…”

miR-9-5p, miR-675-5p and miR-138-5p Damages the Strontium and LRP5-Mediated Skeletal Cell Proliferation, Differentiation, and Adhesion

“…MC3T3-E1 cells (4 × 10 5 cells/well) in the six-well plates were treated with strontium ranelate six hours after seeding. We detected the miRNAs that were related to bone formation according to previous studies [ 12 , 13 , 14 , 15 , 16 , 17 , 18 ]. Results showed the down-regulation of miR-9-5p upon strontium ranelate treatment but there were no significant changes ( Figure 1 A).…”