BMP2诱导C3H10细胞成骨分化的TGFβ-Ⅰ型受体的体外分析

Analyses of TypeⅠTGF-β Receptors in BMP2 Induced Osteogenic Differentiation of C3H10 Cell in Vitro

筛选和分析与BMP2诱导间充质干细胞C3H10成骨分化有关的TGF-βⅠ型受体。利用显性负性突变型TGF-βⅠ型受体竞争抑制配体功能的特性,运用碱性磷酸酶定量测定、Real time PCR等方法,初步筛选出可能与BMP2诱导间充质干细胞C3H10成骨分化有关的的TGF-βⅠ型受体,随后运用RNA干扰的方法抑制相应TGF-βⅠ型受体的表达,进一步证实相关TGF-βⅠ型受体与BMP2发挥诱导成骨活性的关系。结果证实,显性负性突变的ALK3和ALK6能够抑制BMP2诱导的C3H10细胞成骨分化;RNA干扰抑制ALK3或(和)ALK6表达后,BMP2诱导C3H10细胞向成骨分化的趋势受到抑制。因此,ALK3和ALK6是与BMP2诱导C3H10细胞成骨分化有关的TGF-βⅠ型受体。

It was to analysis the type Ⅰ TGF-β receptors in BMP2 induced osteogenic differentiation. The type Ⅰ TGF-β receptors involved in BMP2 induced osteogenic differentiation of C3H10 cell was identified by using ALP quantitative assay and Real time PCR. The expression of BMP2 related type Ⅰ TGF-β receptors was knock down by RNA interference, to confirm the relationship between osteogenie activity of BMP2 and type Ⅰ TGF-β receptors. Results showed that dominant negative forms of ALK3 and ALK6 can inhibit BMP2 induced osteogenic differentiation of C3H10 cell,and ALK3/ALK6 knock-down by RNAi can inhibit BMP2 induced osteogenic differentiation of C3H10 cell. It can prove that ALK3 and ALK6 are the type Ⅰ TGF-β receptors that can involve in BMP2 induced osteogenie differentiation of C3H10 cell.