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Hey1 Gene Is Involved in Regulating BMP9-induced Osteogenic Differentiation of C3H10T1/2 Cells |
TANG Da-gang, WANG Miao, ZHANG Yan-liang, WANG Xiao-lin, LI Gui-qiang, LUO Xiao-ji |
Orthopedic Department, the First Affiliate Hospital of Chongqing Medical University, Chongqing 400016, China |
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Abstract Objective: To analysis the effect of Hey1 gene on BMP9-induced osteogenic differentiation of mesenchymal stem cells C3H10T1/2. Methods: Assemble lentivirus LV5-GFP, LV5-BMP9 and LV5-Hey1, the expression of GFP, BMP9 and Hey1 were detected by RT-PCR and Western blot in C3H10T1/2 cells. Then C3H10T1/2 cells was infected with LV5-GFP, LV5-Hey1 and/or LV5-BMP9 respectively, the early osteogenic marker ALP activity was detected by quantitative assay, the later osteogenic marker calcium deposition was detected by Alizarin Red S staining, the cell proliferation and cycles regulated by BMP9 were detected by MTT and flow cytometry assay. Results: LV5-GFP, LV5-BMP9 and LV5-Hey1 promoted the expression of GFP, BMP9 and Hey1. LV5-Hey1 can increase the early and later osteogenic differentiation of C3H10T1/2 cells induced by BMP9, and promote the cell proliferation in the early stage, but inhibit cell cycles in the later stage regulated by BMP9. Conclusion: Hey1 gene can influence BMP9-induced osteogenic differentiation and BMP9-regulated cell proliferation and cell cycles of C3H10T1/2 cells.
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Received: 26 February 2015
Published: 25 June 2015
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