Expression Of Relative Gene In Osteoblasts Differentiation

Objective:Bone formation is a complex process, in which the mesenchymal cells differentiate into osteoblasts, while secret extracellular matrix. With the calcification of the extracellular matrix and further mineralization of osteoblasts, osteoblasts are embedded in the bone and become bone cells. Osteogenic differentiation of mesenchymal cells can be summarzied briefly as the following stages:mesenchymal stem cells→bone progenitor cells→osteoblast precursor cells→mature osteoblasts→bone cells.It has been proven, mature osteoblasts secrete the DMP1, ALP, PHEX, Sclerostin adjusted to the entire process of osteogenic differentiation. However, as osteoblasts differentiating into bone cells, this literature has not yet give a clear answer to expiaining what kind of changes into the regulation of bone differentiation process.In this study, we conducted osteogenic induction experiments with MC3T3-E1cell lines. Using real-time quantitative RT-PCR method, we study DMP1mRNA, ALP mRNA in PHEX mRNA in SOST mRNA expression changes in the process of osteogenic differentiation of osteoblasts, to provide a theoretical basis for the further elucidate osteoblast differentiation and regulation mechanism.Methods:MC3T3-E1cells were induced for0days,7days,14days,21days,35days with osteogenic inductor. We observed cells mineralized nodule formation at each point in time, then we detected the DMP1, ALP, PHEX SOST gene mRNA expression of various time points, using QIAGEN kit willtotal to extract total RNA, which was reversely transcribed into cDNA, and then using DMP1, ALP, PHEX, SOST primers to run RFP-PCR.Results:As time increases that MC3T3-E1cells differentiated into osteocytes, cell number growth trend slowed down, and mineralized nodules gradually became larger, the number of which gradually increased.As time increases that MC3T3-E1cells differentiated into osteocytes, the expression ofDMP1mRNA, PHEX mRNA SOST mRNA is gradually increased, at the time of35days to reach the highest, wherein the increased expression of the SOST mRNA amount maximum; ALP mRNA expression gradually decreased, at the time of35days to reach the minimum.Conclusion:1. Under the action of osteogenic liquid, osteoblast differentiation to bone cells showing characteristics of bone cell gene expression.2. During osteogenic differentiation, a significant change in the amount of gene expression associated with osteogenesis osteoblasts has also changed its regulatory role of the bone process. The DMP mRNA, PHEX mRNA SOST mRNA expression gradually increased, ALP mRNA expression decreased gradually.