组蛋白去甲基化酶JMJD3对骨髓基质干细胞干性特征及体外成骨分化的影响  

Effects of JMJD3 on stemness and osteogenic differentiation of bone marrow stromal stem cells

作　　者：董伟杰[1] 戈杰[1] 张平[1] 傅瑜[1] 江宏兵[1] 

机构地区：[1]南京医科大学口腔医学研究所.附属口腔医院口腔颌面外科,江苏南京210029

出　　处：《口腔生物医学》2014年第1期6-10,共5页Oral Biomedicine

基　　金：国家自然科学基金(81070810);江苏高校优势学科建设工程资助项目(2011-137)

摘　　要：目的:探讨组蛋白去甲基化酶JMJD3对骨髓基质干细胞(bone marrow stromal cells,BMSCs)体外多潜能性及成骨分化的生物学调控作用。方法:用离心和贴壁培养相结合方法分离培养大鼠四肢骨BMSCs,分别用含JMJD3-shRNA的绿色荧光蛋白(GFP)慢病毒干扰载体(实验组)及含无关序列的GFP慢病毒载体(对照组)转染BMSCs,Western blot方法检测两组BMSCs中组蛋白H3第27位赖氨酸上三甲基化(H3K27me3)及干细胞多潜能转录因子Oct4、Nanog、Sox2的表达,并比较两组细胞的克隆形成率;实时定量RT-PCR、Western blot、茜素红染色检测两组细胞的体外成骨分化能力。结果:与对照组相比,实验组BMSCs的H3K27me3表达水平升高,具有更强的克隆形成能力,且表达更强的Oct4、Nanog、Sox2等多潜能因子;体外成骨诱导7 d后,实验组BMSCs中RUNX2等成骨分化基因表达下降,诱导14 d后,钙结节形成较对照组少。结论:抑制JMJD3表达可增强BMSCs的干性特征,抑制其成骨分化。Objective：To investigate the role of the histone demethylases JMJD 3 in regulating the pluripotency and the osteogenic differentiation of the bone marrow stromal cells （BMSCs）in vitro.Methods：Gradient centrifugation and adherent culture were used to isolate the BMSCs from rat tibia, and the lenti-virus of the JMJD3-shRNA and GFP （as the control） were constructed.The expression of H3K27me3 and the pluripotent transcription factors （ Oct4/Nanog/Sox2） were detected by Western blot .The colony-forming effi-ciency was compared between JMJD 3-shRNA group and GFP group .The different osteogenic ability of the JMJD 3-shRNA group and the control group were analyzed by real-time RT-PCR, Western blot and Alizarin red staining after osteogenic induction .Results：Com-pared with the control, the expression of H3K27me3 was higher in JMJD3-shRNA group.The JMJD3 shRNA group possessed stronger colony-forming ability, expressed higher stemness markers （Oct4/Nanog/Sox2）, compared with the control group .After 7-day osteo-genic induction in vitro , the BMSCs′osteogenic ability declined in JMJD 3-shRNA group .After 14-day osteogenic induction , the calci-um nodules of JMJD3-shRNA group were less than that of the control group .Conclusions：Inhibition of JMJD3 can enhance BMSCs′stemness, and suppress their osteogenic differentiation .

关 键 词：组蛋白去甲基化酶 JMJD3 表观遗传修饰 干性 成骨诱导 JMJD3 

分 类 号：R780.2[医药卫生—口腔医学]