miR-150在骨髓间充质干细胞分化中的作用及其可能的机制  

Role of miR-150 in the Differentiation of Bone Marrow Mesenchymal Stem Cells and its Possible Mechanism

作　　者：郭伟壮[1] 兰涛 杨欣建[1] 田长庆[1] GUO Weizhuang;LAN Tao;YANG Xin-jian(Spine Surgery,Shenzhen Second People’s Hospital,Shenzhen,Guangdong,518000,China)

机构地区：[1]深圳市第二人民医院脊柱外科

出　　处：《黑龙江医学》2019年第10期1143-1146,共4页Heilongjiang Medical Journal

摘　　要：目的采用去卵巢法建立骨质疏松模型,并研究模型中miR-150的表达情况及其影响骨质疏松过程的可能机制。方法选取C57BL/6J采用卵巢摘除法建立骨质疏松模型,micro CT检测模型中骨质情况,并分离BMSCs细胞,流式细胞仪检测BMSCs表面抗原检测结果,MTT生长曲线和克隆形成情况鉴定BMSCs细胞增殖情况,RT-PCR检测模型中miR-150、RUNX-2的表达情况,转染miR-150 mimics和inhibitor后RT-PCR和Western Blot检测细胞中RUNX-2的表达情况,成骨诱导及碱性磷酸酶(ALP)染色和茜红素检测不同处理组成骨能力变化情况。结果处理2个月后实验组骨小梁明显减少,稀疏,断裂,骨含量下降;BMSCs克隆数及细胞增殖能力6 d、8 d后显著降低,miR-150内源性表达异常升高,两组BMSCs表型CD29、CD44阳性,CD34、CD45阴性。模型建立成功后,实验组中RUNX-2表达显著降低,inhibitor抑制miR-150表达后,RUNX-2表达升高,miR-150 mimics组结果变化趋势与miR-150 inhibitor组相反。与对照组相比,mimics组ALP染色结果变浅,表达降低,而inhibitor组染色变深,表达升高。结论在成骨诱导过程中,miR-150可作为成骨负性调控因子,通过影响RUNX-2的表达影响成骨分化。Objective To establish a model of osteoporosis by ovariectomy,and to study the expression of mi R-150 in the modeland its possible mechanisms affecting the osteoporosis process.MethodsC57 BL/6 J was used to establish osteoporosis model by ovarianablation.The bone condition was detected by micro CT,and BMSCs cells were isolated.The surface antigen detection results of BMSCswere detected by flow cytometry.The growth curve and clone formation of BMSCs were used to identify the proliferation of BMSCs.In thecase of RT-PCR,the expression of mi R-150 and RUNX-2 in the model was detected.After transfection of mi R-150 mimics and inhib-itor,RT-PCR and Western Blot were used to detect the expression of RUNX-2 in cells,osteogenic induction and alkaline phosphoricacid.Enzyme(ALP)staining and lycopene detection varied the composition of bone composition.ResultsAfter 2 months of treatment,the trabecular bone of the experimental group significantly reduced,scattered,fractured and decreased.The number of clones and cellproliferation of BMSCs decreased significantly after 6 days and 8 days,and the endogenous expression of mi R-150 was abnormally in-creased.BMSCs were positive for CD29 and CD44,and negative for CD34 and CD45.After successful model establishment,the ex-pression of RUNX-2 was significantly decreased in the experimental group.The expression of RUNX-2 increased after the inhibitor in-hibited the expression of mi R-150,and the change trend of the mi R-150 mimics group was opposite to that of the mi R-150 inhibitorgroup.Compared with the control group,the ALP staining results in the mimics group were lighter and the expression decreased,whilethe inhibitor group was darker and the expression increased.ConclusionIn the process of osteogenic induction,mi R-150 can be usedas a negative osteogenic regulator to affect osteogenic differentiation by affecting the expression of RUNX-2.

关 键 词：RUNX-2 miR-150 BMSCS 

分 类 号：R734.2[医药卫生—肿瘤]