过表达miR-30a抑制BMP9诱导C2C12细胞的成骨分化

基础医学与临床 ›› 2016, Vol. 36 ›› Issue (3): 364-369.

过表达miR-30a抑制BMP9诱导C2C12细胞的成骨分化

李宝林¹,施琼²,邓正华¹,温先勇¹,彭瑛¹,唐敏¹,刘靳波¹

1. 泸州医学院附属医院
2. 重庆医科大学检验医学院

收稿日期:2015-09-10 修回日期:2015-11-30 出版日期:2016-03-05 发布日期:2016-02-22
通讯作者: 刘靳波 E-mail:liujb7203@163.com
基金资助:
2014年四川省科学技术厅科研专项基金

miR-30a over-expression inhibits osteogenic differentiation induced by BMP9 in C2C12 cells

Received:2015-09-10 Revised:2015-11-30 Online:2016-03-05 Published:2016-02-22

摘要/Abstract

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。

关键词: miR-30a, 骨形成蛋白9, C2C12, 成骨分化

Abstract: 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-induced osteogenic differentiation of C2C12 cells were detected by alkaline phosphatase(ALP)，osteocalcin(OCN) and calcium deposition. MTT and cell cycle analysis were used to reveale cell proliferation. Results The expression of only one miR-30 family member, miR-30a, first decreased and then increased during BMP9-induced osteogenic differentiation of C2C12 cells.Over-expression of miR-30a ，which targets on Runx2，led to expression of an early osteogenic marker and a reduction in ALP expression. In addition, we observed decreases in the expression of later osteogenic markers Osteocalcin，as well as calcium deposition. Conclusions BMP9-induced osteogenic differentiation is partially inhibited by miR-30a in mesenchymal stem cell line C2C12.

Key words: miR-30a , Bone morphogenetic protein 9, C2C12, osteogenic differentiation

中图分类号:

R273

李宝林施琼邓正华温先勇彭瑛唐敏刘靳波. 过表达miR-30a抑制BMP9诱导C2C12细胞的成骨分化[J]. 基础医学与临床, 2016, 36(3): 364-369.

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