miR-1908过表达的人脂源性多能干细胞基因表达谱分析

Gene expression proles of miR - 1908 over - expressed human multipotent adipose - derived stem cells by cDNA microarrays

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—timePCR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。

Objective To explore the underlying molecular basis of hsa - miR - 1908 in obesity, and to exami- ne human muhipotent adiposederived 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 bMADS 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 （18/123,14.63o/0）, 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.