再生障碍性贫血骨髓间充质干细胞干细胞相关基因和microRNA表达谱.DOC

再生障碍性贫血患儿骨髓MSCs基因 和miRNA表达谱对照研究 李福兴 蒋莎义 谢晓恬1 周吉吉1 石苇1 邵越霞1 周晓迅1 曹萍1 基金项目：上海市卫生局资助项目（2006091） 1作者单位：200065 上海，同济大学附属同济医院 儿科 2通讯作者：蒋莎义 上海交通大学附属儿童医院 血液科 联系地址：上海市北京西路1400弄24号 上海市儿童医院血液科 联系电话：再生障碍性贫血患儿骨髓MSCs基因 和miRNA表达谱对照研究 【摘要】目的　我们前期实验分离培养再生障碍性贫血患儿骨髓间充质干细胞，发现其增殖能力减弱，提示间充质干细胞存在异常。本实验拟用干细胞功能分类基因芯片与miRNA芯片对照分析，进一步研究骨髓间充质干细胞在再生障碍性贫血发病中的作用。方法　选取再障和正常对照各10例患儿，体外分离、培养骨髓间充质干细胞，并选取两组各2例的P3代骨髓间充质干细胞。利用干细胞功能分类基因芯片和miRNA芯片检测再障与正常对照的差异表达基因和micro-RNA。通过差异基因和差异micro-RNA比对，挑选出FGF2基因和其对应的hsa-miR-107，行Real time PCR进一步验证。结果　与正常对照比较，差异基因32个；差异表达miRNA共62个，2倍以上上调的52个，2倍以上下调的10个。基于生物信息学预测的方法，将差异基因与差异miRNA比对，筛选出hsa-miR-107和其调节基因FGF2，实时定量PCR验证FGF2表达下降和hsa-miR-107高表达。 结论 FGF2和其对应的microRNA hsa-miR-107表达异常的骨髓间充质干细胞可能在再生障碍性贫血患儿的发病中起重要作用。 【关键词】再生障碍性贫血 骨髓间充质干细胞 microRNA芯片 hsa-miR-107 The profiles of microRNA and stem cell related gene in bone marrow mesenchymal stem cells from children with aplastic anemia LI Fu-xing, JIANG Sha-yi , XIE Xiao-tian, ZHOU Ji-ji, SHI Wei, SHAO Yue-xia, ZHOU Xiao-xun, Cao Ping, (1 Department of Pediatrics, Shanghai Tong Ji Hospital, Tong Ji University, Shanghai, China; 2 Department of Hematology, Children?s Hospital of Shanghai, Shanghai Jiao Tong University, Shanghai, China) Abstract: Objectives Isolation and culture of mesenchymal stem cells (MSCs) from bone marrow in children with aplastic anemia in our previous experiments revealed that the proliferation ability of MSCs was reduced. This experiment is a further study of bone marrow mesenchymal stem cells in the pathogenesis of aplastic anemia by analysis of gene chip and miRNA chip. Methods MSCs isolated from the bone marrow of aplastic anemia children and control group were cultured in vitro. Gene expression profiling and expression of microRNA spectrum were examined on P3 cells using functional classification microarray and miRNA chip respectively. Comparing the microRNAs expression profile and stem cell related gene expression profile, all the significantly difference microRNAs and their target g