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| Role of Notch Signaling Pathway in Bone Morphogenetic Protein 9-induced Ostegenic Differentiation of Marrow-derived Mesenchymal Stem Cells and Its Mechanism |
| XIE Jia-ying, XU Wen-chun, XU Dao-jing, ZHANG Xiao-yan, TANG Min |
| Key Laboratory of Clinical Laboratory Diagnostics of Ministry of Education, Faculty of laboratory Medicine, Chongqing Medicine University, Chongqing 400016, China |
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Abstract Objective: To study the role of Notch signaling pathway in bone morphogentic protein 9(BMP9) induced osteogenic diffierentiaion of marrow-derived mesenchymal stem cells and its mechanism. Method: After treatment C2C12 cells with recombinant adenovirus GFP and BMP9 (AdGFP/AdBMP9), the early osteogenic marker Alkaline phosphatase (ALP) activity was detected by staining assay, later osteogenic marker calcium deposition was determined by Alizarin Red S staining. Effects of Notch signaling pathway on ostegenic differentiation was detected by cell-density assay and Notch signaling pathway inhibitor (DAPT). The receptor of BMP9, Smad1/5/8 and some transcription factors were detected in order to study the its mechanism. Results: AdBMP9 is not only increased the activity of ALP on days 3, 5, 7 matrix mineralization on 7d and 14d in a dose-dependent manner. Notch specific inhibitor DAPT decreased ALP activity and the expression of OCN. Furthermore, DAPT markedly change expression level of ALK2 and ostegenic differentiation marker Runx2 and Col1a-α induced by BMP9 of C3H10T1/2. DAPT did not change total protein level of Smad1/5/8, however, DAPT increased the phosphorlated form of Smad1/5/8. Conclusion: Notch signaling pathway can enhance BMP9-induced osteogenic differenitiation of marrow-derived mesenchymal stem cells possibly by up-regulating the expression of its ligands and receptors through BMP9, Notch signaling pathway feedback up-regulates BMP9 signaling, thus leading to osteogensis.
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Received: 17 July 2012
Published: 25 November 2012
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