ZHANG Xiao, XU Daojing, LIN Liangbo, et al. Mechanism of osteogenic differentiation for mesenchymal stem cells induced by BMP9 and Wnt3a in vitro[J]. Journal of Chongqing Medical University, 2012,37(7):565-569.
ZHANG Xiao, XU Daojing, LIN Liangbo, et al. Mechanism of osteogenic differentiation for mesenchymal stem cells induced by BMP9 and Wnt3a in vitro[J]. Journal of Chongqing Medical University, 2012,37(7):565-569.DOI:
目的:探讨经典Wnt信号通路与骨形态发生蛋白9(Bone morphogenetic protein 9,BMP9)诱导间充质干细胞(Mes-enchymal stem cells,MSCs)成骨的相互影响及分子机制。方法:以小鼠骨髓来源MSCs C3H10T1/2为目的细胞,用Wnt3a上调经典Wnt信号通路,联合BMP9作用于细胞,通过碱性磷酸酶(Alkaline phosphatase,ALP)活性测定、钙盐沉积实验、real time PCR、免疫细胞化学染色等方法观测BMP9与经典Wnt信号通路联合后对MSCs成骨分化的影响。结果:Wnt3a明显促进了BMP9诱导的ALP活性的增加[F=264.962,P=0.000;t(BMP9+Wnt3a)-BMP9=7.19,P=0.000;t(BMP9+Wnt3a)-Wnt3a=16.36,P=0.000],同时也明显增加了成骨标志物骨钙蛋白(Osteocalcin,OC)[F=3 920.102,P=0.000;t(BMP9+Wnt3a)-BMP9=39.10,P=0.000;t(BMP9+Wnt3a)-Wnt3a=62.56,P=0.000]和骨桥蛋白(Osteopontin,OPN)[F=1 935.824,P=0.000;t(BMP9+Wnt3a)-BMP9=40.88,P=0.000;t(BMP9+Wnt3a)-Wnt3a=56.42,P=0.000]的表达以及Runx2的mR-NA的表达[F=3 635.980,P=0.000;t(BMP9+Wnt3a)-BMP9=79.37,P=0.000;t(BMP9+Wnt3a)-Wnt3a=86.96,P=0.000],并且还促进了钙盐沉积。结论:Wnt3a与BMP9相互协同诱导MSCs的骨向分化,其中成骨转录因子Runx2可能作为二者的交汇点起了重要作用。
Abstract
Objective:To analyze the molecular mechanism of osteogenic differentiation of mesenchymal stem cells(MSCs) induced by canonical Wnt3a signal pathway and bone morphogenetic protein 9(BMP9). Methods:C3H10T1/2 cells were infected by recombinant adenovirus expressing BMP9 and Wnt3a,and then alkaline phosphatase(ALP),the early osteogenic marker,was detected by quantita-tive assay. Osteogenic differentiation marker at later stage,calcium deposition was determined by Alizarin Red S staining. The protein levels of osteocalcin(OC) and osteopontin(OPN) were detected by immunocytochemical staining. The expression levels of OC,OPN and Runx2 were analyzed by real time PCR. Results:Wnt3a enhanced the ability of BMP9-induced ALP activity(F=264.962,P=0.000;t(BMP9+Wnt3a)-BMP9=7.19,P=0.000;t(BMP9+Wnt3a)-Wnt3a=16.36,P=0.000) and increased expressions of OC(F=3 920.102,P=0.000;t(BMP9+Wnt3a)-BMP9=39.10,P=0.000;t(BMP9+Wnt3a)-Wnt3a=62.56,P=0.000) and OPN(F=1 935.824,P=0.000;t(BMP9+Wnt3a)-BMP9=40.88,P=0.000;t(BMP9+Wnt3a)-Wnt3a=56.42,P=0.000),with increase of mineral calcium deposition in vitro. Furthermore,Wnt3a increased the mRNA level of Runx2(F=3 635.980,P=0.000;t(BMP9+Wnt3a)-BMP9=79.37,P=0.000;t(BMP9+Wnt3a)-Wnt3a=86.96,P=0.000). Conclusion:Wnt3a,possibly through Runx2,acts synergistically on BMP9 induced osteogenic differentiation of MSCs.
关键词
骨形态发生蛋白9Wnt3a间充质干细胞骨向分化
Keywords
bone morphogenetic protein 9Wnt3amesenchymal stem cellsoseteogenic differentiation