Runx3敲减抑制BMP9诱导小鼠胚胎成纤维细胞iMEFs向成骨分化

基础医学与临床 ›› 2016, Vol. 36 ›› Issue (2): 173-178.

Runx3敲减抑制BMP9诱导小鼠胚胎成纤维细胞iMEFs向成骨分化

王玉凤¹,冯巧玲²,吉彩霞²,刘晓骅²,罗进勇¹

1. 重庆医科大学
2. 重庆医科大学检验医学院

收稿日期:2015-04-30 修回日期:2015-09-28 出版日期:2016-02-05 发布日期:2016-01-21
通讯作者: 罗进勇 E-mail:luojinyong@sina.com
基金资助:
BMP9/PI3K/AKT信号途径调控间充质干细胞成骨分化;重庆市基础与前沿研究计划项目

Knocking down Runx3 inhibits BMP9-induced osteogenic differentiation of iMEFs cells

Received:2015-04-30 Revised:2015-09-28 Online:2016-02-05 Published:2016-01-21

摘要/Abstract

摘要： 目的研究和确认Runx3敲减对BMP9诱导的小鼠胚胎成纤维细胞iMEFs成骨分化的作用。方法BMP9腺病毒感染iMEFs,用Western blot检测内源性Runx3蛋白的表达。Runx3干扰腺病毒ad-siRunx3和BMP9条件培养基共同处理iMEFs，用碱性磷酸（ALP）染色和活性定量测定，检测成骨早期指标ALP；用茜素红S染色，检测成骨晚期指标钙盐沉积；用RT-PCR检测成骨相关基因Id1、Id2、Id3和Runx2，用 Western blot检测成骨相关蛋白Dlx5；最后，用SBE荧光素酶报告质粒，检测smad1/5/8的转录活性。结果：BMP9可以使Runx3表达降低（P<0.05）；干扰Runx3可以抑制早期成骨指标ALP活性（P<0.05）和晚期成骨指标钙盐沉积；ad-siRunx3抑制BMP9诱导的成骨相关转录因子Id1、Id2、Id3和Runx2基因的表达（P<0.05）及DLX5蛋白的表达(P<0.05)。结论敲减Runx3可以抑制BMP9诱导小鼠胚胎成纤维细胞向成骨分化。

关键词: iMEFs, Runx3 , BMP9 , 成骨分化

Abstract: Objective： To analyse knocking down Runx3 on BMP9-induced osteogenic differenciation of iMEFs. Methods：iMEFs were infected with ad-BMP9 and then the expression of endogenous Runx3 was determined by RT-PCR and Western blot. iMEFs were treated with ad-siRunx3 and BMP9,then the alkaline phosphatase( ALP) activity was detected by quantitative and staining assay; calcium deposition was detected by Alizarin Red S staining ; the gene expression levels of Id1,Id2,Id3 and Runx2 were determined by RT-PCR and the protein expression level of DLX5 was determined by western blot. iMEFs were transfected with p(12SBE)- Luc and detected by quantitative. Results BMP9 can inhibit the expression of Runx3.Ad-siRunx3 can inhibit the expression of ALP, which is early osteogenic marker, and the expression of calcium deposition, which is late osteogenic marker . Ad-siRunx3 can inhibit the expression of Id1,Id2, Id3, Runx2 and DLX5 induced by BMP9. Conclusion Knocking down Runx3 can inhibit the BMP9-induced Osteogenic differentiation of iMEFs.

Key words: iMEFs , Runx3 , BMP9 , osteogenic differentiation

中图分类号:

Q254

王玉凤冯巧玲吉彩霞刘晓骅罗进勇. Runx3敲减抑制BMP9诱导小鼠胚胎成纤维细胞iMEFs向成骨分化[J]. 基础医学与临床, 2016, 36(2): 173-178.

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