Effect of activator protein 2α on BMP9-induced osteogenesis differentiation of C3H10 and its mechanism
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Effect of activator protein 2α on BMP9-induced osteogenesis differentiation of C3H10 and its mechanism
Journal of Chongqing Medical UniversityVol. 40, Issue 2, Pages: 181-185(2015)
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Published:2015,
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Effect of activator protein 2α on BMP9-induced osteogenesis differentiation of C3H10 and its mechanism. [J]. Journal of Chongqing Medical University 40(2):181-185(2015)
DOI:
Effect of activator protein 2α on BMP9-induced osteogenesis differentiation of C3H10 and its mechanism. [J]. Journal of Chongqing Medical University 40(2):181-185(2015)DOI:
Effect of activator protein 2α on BMP9-induced osteogenesis differentiation of C3H10 and its mechanism
目的:研究转录蛋白2(activator protein 2,AP2)α在骨形态发生蛋白(bone morphogenetic protein,BMP)9诱导小鼠间充质干细胞C3H10成骨分化中的作用及可能的分子机制。方法:重组腺病毒 Ad-BMP9感染小鼠C3H10细胞,显性负性突变体AP2α-△bHLH和AP2α-△TAD抑制 AP2α转录活性,通过RT-PCR、real-time PCR检测AP2α、BMP9及成骨相关基因的表达水平,Western blot检测骨桥蛋白(osteopontin,OPN)表达,碱性磷酸酶(alkaline phosphatase,ALP)活性测定、茜红素染色实验观测C3H10成骨分化。结果:BMP9具有强促成骨分化的作用,对AP2α的表达水平无影响,AP2α显性负性突变体可抑制BMP9诱导的成骨基因骨钙蛋白(osteocalcin,OC)、OPN及骨保护素(osteoprotegerin,OPG)表达的增高(OC:F=56.929,P=0.000;OPN:F=54.789,P=0.000;OPG:F=159.451,P=0.000),ALP活性(F=69.776,P=0.000)及红色钙盐沉积减少。结论:BMP9可能通过影响AP2α的转录活性调控成骨分化进程。
Abstract
Objective:To research the effect of activator protein 2α on the process of bone morphogenetic protein 9(BMP9)-induced osteogenesis differentiation of mouse mesenchymal stem cells(C3H10) and its mechanism. Methods:C3H10 cells were infected with Ad-BMP9. Adenovirus-mediated dominant negative mutants Ad-dnAP2α-△bHLH and Ad-dnAP2α-△TAD were used to inhibit AP2α transcriptional activity. RT-PCR,real-time PCR and Western blot methods were used to detect the expression of AP2α,BMP9 and osteogenetic related markers. Alkaline phosphatase reading/staining and alizarin red staining were performed to detect the status of osteogenetic differentiation. Results:Over-expression of BMP9 could accelerate osteogenesis differentiation but did not affect the expression of AP2α. AP2α dominant negative mutants could inhibit the BMP9-induced increase of osteogenesis related markers osteocalcin(OC),osteopontin(OPN)and osteoprotegerin(OPG)expression(OC:F=56.929,P=0.000;OPN:F=54.789,P=0.000;OPG:F=159.451,P=0.000)and reduce the alkaline phosphatase activity(F=69.776,P=0.000) and red staining of calcium salt deposi-tion. Conclusion:AP2α transcription activity might regulate the process of BMP9-induced osteogenesis differentiation.
关键词
转录蛋白AP2α显性负性突变体骨形态发生蛋白9成骨分化
Keywords
activator protein 2αdominant negative mutantbone morphogenetic protein 9osteogenesis differentiation
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