摘要目的 研究miR-1908对脂源性多能干细胞基因表达谱的影响,并探讨其在肥胖发生过程中的可能作用.方法 构建miR-1908过表达的脂源性多能干细胞,利用基因表达谱芯片技术筛选得到与阴性对照组差异表达的基因.挑选3个差异表达基因,利用Real-time PCR技术验证miRNA芯片检测结果.结果 基因表达谱芯片结果显示,差异表达的123个基因中,2倍以上上调的基因有69个,下调的有54个,差异均有统计学意义(P均＜0.05).将这些基因按照功能分类,分为15组:高分子配合物亚基基因(18/123,14.63％),RNA结合相关基因(16/123,13.01％),凋亡相关基因(14/123,11.38％),细胞程序性死亡相关基因(14/123,11.38％),磷酸酶活性相关基因(9/123,7.32％),蛋白结构域结合相关基因(10/123,8.13％),蛋白磷酸酶活性相关基因(8/123,6.50％),分子成分分解相关基因(5/123,4.07％),肾脏发育相关基因(6/123,4.88％),RNA聚合酶Ⅱ转录启动子基因(9/123,7.32％),泌尿生殖系统发育相关基因(6/123,4.88％),依赖DNA的转录基因(9/123,7.32％),RNA合成相关基因(9/123,7.32％),Ⅰ型干扰素合成过程相关基因(2/123,1.63％)及Ⅰ型干扰素合成产物相关基因(2/123,1.63％).结论 miR-1908可能在肥胖发生的过程中发挥调控作用.在下调表达的基因中验证了miR-1908可能直接作用的靶基因,为研究miR-1908在肥胖发生中的作用奠定了基础,同时也为miRNA的生物学功能及其作用机制的研究提供了参考.

abstractsObjective To explore the underlying molecular basis of hsa-miR-1908 in obesity,and to examine human multipotent adipose-derived stem (hMADS) cell genes differentially expressed in hsa-miR-1908 overexpressed hMADS cell by using cDNA microarrays.Methods A lentiviral vector containing miR-1908 with high overexpression in the infected hMADS cells was constructed.Gene expression proles of hsa-miR-1908 over-expressed human multipotent adipose-derived stem cells were obtained by cDNA microarrays.Three of the differentially regulated genes identified with cDNA microarray analysis were randomly selected for validation analysis by Real-time quantitative PCR.Results cDNA microarrays containing 34 127 genes were used to investigate gene expression of hMADS cells.The analysis of gene expression proles indicated that 69 genes were up-regulated and 54 genes were down-regulated in hsa-miR-1908 over-expressed hMADS cells.Based on their molecular functions,these genes were classified into 15 different groups,including macromolecular complex subunit organization (18/123,14.63％),RNA binding (16/123,13.01％),phosphatase activity (9/123,7.32％),protein domain specific binding (10/123,8.13 ％),phosphoprotein phosphatase activity (8/123,6.50％),cellular component disassembly (5/123,4.07％),kidney development (6/123,4.88％),transcription from RNA polymerase Ⅱ promoter (9/123,7.32％),urogenital system development (6/123,4.88％),transcription DNA-dependent (9/123,7.32％),RNA biosynthetic process (9/123,7.32％),apoptosis (14/132,11.38％),programmed cell death (14/123,11.38％),type Ⅰ interferon biosynthetic process (2/123,1.63％),and type Ⅰ interferon production (2/123,1.63％).Conclusions The present study provides a new view on the study of hsa-miR-1908,unraveling some of the molecular events taking place in hMADS cells,adding candidate genes for future investigation.