摘要
目的探讨miR-30a在BMP9诱导间充质干细胞C2C12成骨分化中的作用。方法 BMP9条件培养基处理间充质干细胞C2C12诱导成骨分化,用real-time PCR连续检测miR-30家族成员的mRNA。用BMP9条件培养基作用Ad-mmu-miR-30a预处理的C2C12细胞,通过ALP染色和活性检测来反映早期成骨指标ALP的表达、分别在mRNA水平和蛋白水平检测中期成骨指标(OCN)的表达、用钙盐沉积来检测晚期成骨分化,用流式细胞术和MTT检测C2C12细胞的周期与增殖。最后探讨miR-30a在BMP9诱导C2C12细胞成骨分化中的作用机制。结果 miR-30家族中,只有miR-30a在BMP9诱导C2C12细胞成骨分化中表达下调,miR-30a过表达后,通过靶基因Runx2使BMP9诱导成骨分化能力减弱,ALP的表达下降(P<0.05),OCN蛋白水平和mRNA水平也都表达下调(P<0.05),钙盐沉积受到抑制。结论 miR-30a可以抑制BMP9诱导C2C12细胞的成骨分化。
Objective To study the role of miR-30a in bone morphogenetic protein 9 （BMP9） induced osteogenic differentiation of mesenchymal stem cell line C2C12. Methods C2C12 cells were treated with BMP9 conditioned medium to induce osteogenic differentiation. Real-time PCR was used to detect the expression of miR-30 family members in BMP9-induced osteogenic differentiation of C2C12 cells. Effects of miR-30a on BMP9-indueed osteo- genic differentiation of C2C12 cells were detected by alkaline phosphatase （ALP）, osteocalcin（OCN） and calcium deposition. MTT and cell cycle analysis were used to demonstrate cell proliferation. Results The expression of on- ly one miR-30 family member, miR-30a, first decreased and then increased during BMP9-indueed osteogenic dif- ferentiation of C2C12 cells. Over-expression of miR-30a,which targets at Runx2 ,led to expression of an early osteo- genic marker and a reduction in ALP expression. In addition, we observed decreases in the expression of later os- teogenic markers Osteocalcin, as well as calcium deposition. Conclusions BMP9-induced osteogenic differentiationis partially inhibited by miR-30a in mesenchymal stem cell line C2C12.
出处
《基础医学与临床》
CSCD
2016年第3期364-369,共6页
Basic and Clinical Medicine
基金
2014年四川省科学技术厅科研专项基金(14JC0801)
