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 ta... More
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.
