Runx1促进BMP9诱导的间充质干细胞MEFs的成骨分化
发布时间:2018-09-07 19:24
【摘要】:目的:研究Runx1在骨形态发生蛋白9(bone morphogenetic proteins 9,BMP9)诱导小鼠胚胎成纤维细胞(mouse embryonic fibroblasts,MEFs)成骨分化中的作用。方法:用BMP9腺病毒感染MEFs细胞,利用RT-PCR和Western blot分别在mRNA和蛋白质水平检测Runx1的内源性表达;构建过表达Runx1的重组腺病毒Ad-Runx1,并在mRNA和蛋白质水平验证Ad-Runx1的效果;用Ad-Runx1和BMP9条件培养基共处理MEFs,检测成骨早期指标碱性磷酸酶(ALP)染色和活性,茜素红S染色检测成骨晚期指标钙盐沉积;RT-PCR和Western blot分别检测成骨关键转录因子Runx2在mRNA和蛋白质水平的变化。结果:Ad-BMP9处理MEFs细胞后,可使Runx1在mRNA和蛋白质水平表达上调;构建的Ad-Runx1处理MEFs后,可使Runx1在mRNA和蛋白质水平表达上调;Ad-Runx1处理BMP9诱导的MEFs细胞后,增强了ALP活性和钙盐沉积以及Runx2在mRNA和蛋白质水平的表达。结论:Runx1可以促进BMP9诱导的间充质干细胞MEFs的成骨分化。
[Abstract]:Aim: to investigate the role of Runx1 in osteogenic differentiation of mouse embryonic fibroblasts (mouse embryonic fibroblasts,MEFs) induced by bone morphogenetic protein 9 (bone morphogenetic proteins 9 (BMP9). Methods: BMP9 adenovirus was used to infect MEFs cells, and RT-PCR and Western blot were used to detect the endogenous expression of Runx1 at the mRNA and protein levels, respectively, and the recombinant adenovirus Ad-Runx1, expressing Runx1 was constructed, and the effect of Ad-Runx1 was verified at mRNA and protein levels. (ALP) staining and activity of alkaline phosphatase (ALP) were detected by co-treatment of MEFs, with Ad-Runx1 and BMP9 conditioned medium in the early stage of osteogenesis. Alizarin red S staining was used to detect the changes of Runx2 in mRNA and protein levels in late osteogenic stage by RT-PCR and Western blot, respectively. Results the expression of Runx1 and protein in MEFs cells were up-regulated after treated with: Ad-BMP9, and Runx1 was up-regulated at mRNA and protein levels in BMP9 induced MEFs cells after MEFs was treated with constructed Ad-Runx1. Enhanced ALP activity, calcium deposition and Runx2 expression at mRNA and protein levels. Conclusion 1: Runx 1 can promote the osteogenic differentiation of mesenchymal stem cells (MEFs) induced by BMP9.
【作者单位】: 重庆医科大学检验医学院重庆医科大学检验医学教育部重点实验室重庆市重点实验室;
【基金】:国家自然科学基金资助项目(31071304,81272006)
【分类号】:R329.2
[Abstract]:Aim: to investigate the role of Runx1 in osteogenic differentiation of mouse embryonic fibroblasts (mouse embryonic fibroblasts,MEFs) induced by bone morphogenetic protein 9 (bone morphogenetic proteins 9 (BMP9). Methods: BMP9 adenovirus was used to infect MEFs cells, and RT-PCR and Western blot were used to detect the endogenous expression of Runx1 at the mRNA and protein levels, respectively, and the recombinant adenovirus Ad-Runx1, expressing Runx1 was constructed, and the effect of Ad-Runx1 was verified at mRNA and protein levels. (ALP) staining and activity of alkaline phosphatase (ALP) were detected by co-treatment of MEFs, with Ad-Runx1 and BMP9 conditioned medium in the early stage of osteogenesis. Alizarin red S staining was used to detect the changes of Runx2 in mRNA and protein levels in late osteogenic stage by RT-PCR and Western blot, respectively. Results the expression of Runx1 and protein in MEFs cells were up-regulated after treated with: Ad-BMP9, and Runx1 was up-regulated at mRNA and protein levels in BMP9 induced MEFs cells after MEFs was treated with constructed Ad-Runx1. Enhanced ALP activity, calcium deposition and Runx2 expression at mRNA and protein levels. Conclusion 1: Runx 1 can promote the osteogenic differentiation of mesenchymal stem cells (MEFs) induced by BMP9.
【作者单位】: 重庆医科大学检验医学院重庆医科大学检验医学教育部重点实验室重庆市重点实验室;
【基金】:国家自然科学基金资助项目(31071304,81272006)
【分类号】:R329.2
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