Ebrahimzadeh-Vesal R, Shokrgozar M A, Nayernia K, Teimoori-Toolabi L, Estiar M A, Miryounesi M, Nourashrafeddin S, Modarressi M H
Faculty of Medicine, Neyshabur University of Medical Science Department of Basic Medical Science Neyshabur Iran.
Pasteur Institute of Iran National Cell Bank of Iran Tehran Iran.
Cell Mol Biol (Noisy-le-grand). 2015 Jul 31;61(3):84-91.
MicroRNAs are new classes of small non—coding regulatory RNAs which control degradation or suppress translation of its target mRNAs by sequence complementarity. Mature microRNAs are enriched in embryonic stem cells and play important roles in controlling stem cell self—renewal as well as control of differentiation. There is significant evidence that microRNAs are involved in the regulation of stem cell differentiation. The male mouse Embryonic Stem Cell line C57BL6/J with normal karyotype 46, XY was used for profiling microRNA expression in undifferentiated mouse embryonic stem cells (mESCs) and mESCs which were differentiated to germ line cells to determine and compare differences in microRNA expression before and after differentiation. Also, testis tissue samples of a 5—day—old mouse and a mature mouse was used as in vivo control. Profiling was performed by quantitative real—time PCR using locked nucleic acid microRNA—specific LNATM—enhanced primers. After data analysis and comparison of results profiled microRNAs expression, three microRNAs, mmu—miR—21, mmu—miR—21* and mmu—miR—16 showed 50.31, 43.76 and 46.77—fold change increase of expression, respectively, in differentiated mESCs in comparison with undifferentiated state with significant p—value (Average p—value p<0.001 for each members of microRNAs). Expression of Let—7 microRNA family increased in differentiated state when compared with undifferentiated mESCs (Average p—value<0.0001 for each members of family). The levels of expression all other profiled microRNAs were significantly higher in undifferentiated in comparison with differentiated mESCs and their expression was down regulated after differentiation. (Average p—value <0.003 for each members of microRNAs).
微小RNA是一类新型的小非编码调节性RNA，其通过序列互补性控制靶mRNA的降解或抑制其翻译。成熟的微小RNA在胚胎干细胞中富集，并在控制干细胞自我更新以及分化过程中发挥重要作用。有大量证据表明微小RNA参与干细胞分化的调控。使用具有正常核型46, XY的雄性小鼠胚胎干细胞系C57BL6/J来分析未分化的小鼠胚胎干细胞（mESC）以及分化为生殖系细胞的mESC中的微小RNA表达，以确定并比较分化前后微小RNA表达的差异。此外，将5日龄小鼠和成年小鼠的睾丸组织样本用作体内对照。使用锁定核酸微小RNA特异性LNATM增强引物通过定量实时PCR进行分析。在对分析的微小RNA表达结果进行数据分析和比较后，与未分化状态相比，三种微小RNA，即mmu-miR-21、mmu-miR-21*和mmu-miR-16在分化的mESC中的表达分别增加了50.31、43.76和46.77倍，p值显著（每种微小RNA成员的平均p值p<0.001）。与未分化的mESC相比，Let-7微小RNA家族在分化状态下表达增加（家族中每个成员的平均p值<0.0001）。与分化的mESC相比，所有其他分析的微小RNA在未分化状态下的表达水平显著更高，并且它们的表达在分化后下调。（每种微小RNA成员的平均p值<0.003）
