MicroRNA-20a通过调节CKIP-1促进小鼠C3H/10T1/2成骨分化

MicroRNA-20a Promotes Osteogenic Differentiation of C3H/10T1/2 Cells through Regulating CKIP-1 Expression

目的:研究microRNA-20a(miR-20a)对小鼠C3H/10T1/2细胞成骨分化作用的影响及其调控机制。方法:对贴壁培养的小鼠C3H/10T1/2细胞成骨诱导不同时间,ALP染色及应用qRT-PCR验证其诱导结果,同时观察MiR-20a在诱导过程中表达变化;细胞转染MiR-20a mimics,CKIP-1 siRNA,在荧光显微镜下观察转染效果,应用qRT-PCR测定过表达及干扰效率;qRT-PCR检测过表达MiR-20a及干扰CKIP-1对细胞成骨分化的影响及过表达MiR-20a对CKIP-1基因表达的影响。结果:成骨标记基因ALP、OSX、OCN、BSP随小鼠C3H/10T1/2细胞诱导过程逐渐升高,且成骨诱导分化过程中MiR-20a表达上调。过表达MiR-20a促进C3H/10T1/2细胞成骨分化,成骨标记基因碱性磷酸酶(ALP)、RUNX2、OSX、0CN、BSP显著高于对照组,并且抑制CKIP-1的表达;干扰CKIP-1能促进细胞成骨标记基因表达。结论:MiR-20a可能通过下调骨形成负调节因子CKIP-1的表达促进C3H/10T1/2细胞成骨分化。

Objective：To investigate the effect of microRNA-20a（MiR-20a） on osteogenic differentiation of mouse C3H/10T1/2 cells and its regulatory mechanism.Methods：Osteogenic differentiation of C3H/10T1/2 cells were identified by ALP staining and qRT-PCR.MiR-20 a mimics and CKIP-1 siRNA were transfected into C3H/10T1/2 cells respectively with lipo3000.The expression of osteoblast marker genes,miR-20 a and CKIP-1 were quantitatively assessed by qRT-PCR.Results：miR-20 a expression was up-regulated during osteoblast differentiation of C3H/10T1/2 cells.Overexpression of miR-20 a promoted osteogenic differentiation.Furthermore,miR-20 a inhibited the expression of bone formation negative regulator CKIP-1.Additionally,CKIP-1 knockdown promoted osteogenic differentiation.Conclusion：MiR-20 a promotes osteogenic differentiation of C3H/10T1/2 cells possibly through inhibiting the expression of CKIP-1.