MiR-21协同BMP9促进间充质干细胞C3H10T1/2成骨分化  

MiR-21 Synergizes with BMP9 in Osteogenic Differentiation During Mesenchymal Stem Cells Line C3H10T1/2

作　　者：宋琪玲[1] 施琼[1] 陈楚[1] 唐祖川 刘红霞[1] 周一青[1] 张汝益[1] 严树涓[1] 翁亚光[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国生物工程杂志》2016年第2期22-29,共8页China Biotechnology

基　　金：国家自然科学基金(NSFC 31200971);教育部博士点基金(20115503110009);重庆市教育委员会科学技术研究项目(KJ120327)资助项目

摘　　要：目的:探讨miR-21与BMP9之间的关系,明确miR-21在BMP9诱导间充质干细胞成骨分化中的作用。方法:(1)Ad-BMP9感染C3H10T1/2细胞,Real-time-PCR检测miR-21表达。RTPCR检测ALP的表达。(2)MiR-21转染C3H10T1/2细胞,Real-time-PCR检测miR-21和BMP9表达。(3)MiR-21和BMP9-CM处理C3H10 T1/2细胞,ALP活性和染色实验检测C3H10 T1/2细胞早期成骨能力。茜素红S染色实验检测钙盐沉积情况。(4)MiR-21和BMP9-CM处理C3H10T1/2细胞,Real-time-PCR检测成骨分化相关因子ALP,OCN的表达。(5)MiR-21和BMP9-CM处理C3H10T1/2细胞,Western blot检测p-Smad1/5蛋白水平的表达。结果:(1)BMP9暂时降低miR-21的表达。MiR-21也可以暂时降低BMP9的表达。(2)MiR-21可以协同BMP9增强ALP和钙盐沉积。(3)MiR-21协同BMP9增加了p-Smad1/5蛋白水平的表达。结论:MiR-21与BMP9存在相互关系,两者可以互相调节表达。MiR-21可以协同BMP9促进间充质干细胞C3H10T1/2细胞成骨分化,这一过程与增强BMP9/Smad信号的激活程度有关。Objective ： （ 1 ） To study if there is a crosstalk between bone morphogenetic protein 9 （ BMP9 ） and microRNA-21 （miR-21） ;（2） to investigate the effect of miR-21 on BMP9-induced osteogenic differentiation of mesenchymal stem cells line C3H10T1/2. Methods： （1） Using Ad-BMP9 infecting C3H10 T1/2 cells, the expression of miR-21 were detected by Real-time-PCR. After transfecting miR-21 mimic 1 day and 5 days, the expression of BMP were tested by Real-time-PCR. （2） In vitro, after transfecting miR-21, BMP9-CM treating C3H10T1/2,ALP staining and ALP activity were detected and the calcium deposition were detected by Alizarin Red S staining. （3） The expression of BMP9 downstream factors ALP and OCN were detected by Real-time-PCR. （4） The protein expression of p-Smadl/5 were detected by Western blot. Results： BMP9 can decrease the expression of miR-21 temporarily and miR-21 also decreases the expression of BMP9 temporarily. After transfecting miR-21 and treating C3H10T1/2 cell with BMP9-CM, the early osteogenic differentiation marker ALP activity and ALP staining and later osteogenic differentiation marker calcium deposition were enhanced. What＇ s more,the expression of ostengenic factors ALP and OCN also increase during miR-21 combined with BMP9-CM. The protein expression of crosswalk between BMP9 p-Smadl/5 were upregulation during the same process. Conclusion： There is a and miR-21. MiR-21 synergizes with BMP9 promoting mesenchymal stem cells C3H10T1/2 osteogenic differentiation by increasing BMP9/Smad signaling pathway activated

关 键 词：MIR-21 骨形成蛋白9 C3H10T1/2细胞 成骨分化 

分 类 号：Q819[生物学—生物工程]