Wnt10b与骨形态发生蛋白9诱导间充质干细胞成骨分化的关系

目的研究Wnt10b与骨形态发生蛋白9(BMP9)诱导间充质干细胞(MSCs)骨向分化的关系,以及相关的分子机制。方法利用PCR、Westernblot、组织化学染色等方法,检测BMP9对Wnt10b表达的影响,以及Wnt10b对BMP9诱导的成骨分化的影响。同时,通过定量PCR、Westernblot、油红O染色、流式细胞术等,分析Wnt10b影响BMP9成骨分化诱导作用的可能机制。结果Wnt10b在C3H10T1/2、C2C12、MEFs和MC3T3-E1细胞中均有表达,BMP9在C3H10T1/2细胞中上调Wnt10b的表达水平。Wnt10b增强BMP9在C3H10T1/2细胞中增加OCN蛋白水平和促进钙盐沉积的能力,沉默Wnt10b减弱BMP9的作用。Wnt10b并不改变BMP9对细胞周期的影响,但能增强BMP9诱导的Smad1/5/8磷酸化,沉默Wnt10b减弱BMP9对Smad1/5/8磷酸化的促进作用。此外,Wnt10b抑制BMP9在C3H10T1/2细胞中诱导的成脂分化,沉默Wnt10b则促进BMP9的成脂分化诱导作用。结论Wnt10b可以促进BMP9诱导的MSCs骨向分化,这种作用可能与增强BMP/Smad信号转导有关。

Relationship between Wnt10b and bone morphogenetic protein-9 induced osteogenic differentiation of mesenchymal stem cells

Aim To study the relationship between Wnt10b and bone morphogenetic protein-9(BMP9)-induced osteogenic differentiation of mesenchymal stem cells (MSCs),and the molecular mechanisms underlying this process.Methods PCR,Western blot and histochemical staining were used to detect the effect of BMP9 on Wnt10b and the effect of Wnt10b on BMP9-induced osteogenic differentiation in MSCs.Meanwhile,real-time PCR,Western blot,oil red O staining,and flow cytometry assay were used to analyze the potential mechanism of Wnt10b affecting the function of BMP9.Results Wnt10b could be detected in C3H10T1/2,C2C12,MEFs and MC3T3-E1 cells.BMP9 up-regulated the expression of Wnt10b in C3H10T1/2 cells.Wnt10b enhanced the capability of BMP9 to increase the level of OCN and mineralization in C3H10T1/2 cells,and silencing Wnt10b attenuated these effects of BMP9.Wnt10b exhibited no substantial effect on cell cycle affected by BMP9,but it enhanced the effect of BMP9 on inducing phosphorylation of Smad1/5/8.While silencing Wnt10b attenuated this effect of BMP9.In addition,Wnt10b inhibited BMP9-induced adipogenic differentiation in C3H10T1/2 cells,and silencing Wnt10b promoted this effect of BMP9.Conclusions Wnt10b can promote BMP9-induced osteogenic differentiation of MSCs,which may be mediated through enhancing BMP/Smad signaling and reducing adipogenic differentiation.