Osterix和核结合因子在成骨样细胞系分化过程中的表达

目的观察小鼠前成骨细胞MC3T3E1的诱导成骨过程中,Osterix(OSX)和核结合因子(Cbfa1)及其两种亚型Cbfa1/P56、Cbfa1/P57表达的变化,了解OSX和Cbfa1表达与成骨细胞分化的关系,探求与OSX变化趋势一致的Cbfa1亚型。方法在小鼠前成骨细胞MC3T3E1培养的不同时间(0、4、10、14、18、22 d),用半定量RT-PCR法检测OSX、Cbfa1、Cbfa1/P56、Cbfa1/P57 mRNA的表达;用Western-blot法检测Cbfa1蛋白的表达;Van Gie-Son苦味酸酸性复红染色法染色细胞Ⅰ型胶原;茜素红染色观察矿化结节形成。结果MC3T3E1在诱导成骨的培养条件下,从培养第18天开始出现矿化,第22天观察到明显的矿化结节形成。OSX mRNA、Cbfa1 mRNA和Cbfa1蛋白的表达量随MC3T3E1的成熟矿化逐渐增高,而Cbfa1/P56 mRNA表达量无变化,但Cbfa1/P57 mRNA在培养第18天和第22天的表达量明显增高。结论在MC3T3E1分化过程中,OSX和Cbfa1的表达量随成骨细胞的成熟逐渐增高。在Cb-fa1的亚型中,Cbfa1/P57的表达与OSX表达趋势一致。

The Expression of Osterix and Cbfa1 in Osteoblasts Differentiation

Objective To observe the gene expression profile of osterix,Cbfa1,Cbfa1/P56,Cbfa1/P57 in MC3T3E1 cells differentiation.Methods The expressions of osterix,Cbfa1,Cbfa1/P56,Cbfa1/P57 mRNA in MC3T3E1 cell line were examined by semiquantitive RT-PCR;the expression of Cbfa1 protein was observed by Western blot.MC3T3E1 cells were stained by Van Gieson and alizarin red method.Results MC3T3E1 cells displayed small mineralization nodules formation on the 18th day,and became more prominent on the 22nd day.The expression of osterix,Cbfa1,Cbfa1/P57 mRNA was gradually increased with the culture time,and the expression of Cbfa1/P56 mRNA was not changed.Conclusion During the differentiation of MC3T3E1 cells,the increased expression of osterix is according with the increased expression of Cbfa1 and Cbfa1/P57.