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The influence of MicroRNA‐150 in Osteoblast Matrix Mineralization
Author(s) -
Dong ChunLing,
Liu HaoZhi,
Zhang ZhenChun,
Zhao HuanLi,
Zhao Hui,
Huang Yan,
Yao JianHua,
Sun TianSheng
Publication year - 2015
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25245
Subject(s) - osteoblast , chemistry , antagomir , mineralization (soil science) , alkaline phosphatase , western blot , microrna , microbiology and biotechnology , type i collagen , cell culture , medicine , endocrinology , in vitro , biology , gene , biochemistry , genetics , organic chemistry , nitrogen , enzyme
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. J. Cell. Biochem. 116: 2970–2979, 2015. © 2015 Wiley Periodicals, Inc.