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目的筛选能有效抑制大鼠骨髓间充质干细胞Mash1基因表达的siRNA序列。方法根据siRNA靶序列设计原则,设计并化学合成针对大鼠Mash1基因编码区的siRNA三对,并以一对无关序列的寡核苷酸作阴性对照。原代培养大鼠骨髓间充质干细胞并分为5组:Mash1-1组,Mash1-2组,Mash1-3组,无关序列组,空白组。用阳离子脂质体LipofectamineTM 2000转染,转染后48h行RT-PCR检测Mash1表达水平的变化。结果转染48h后,与序列3、无关序列、空白组细胞相比,序列1、序列2转染的大鼠骨髓间充质干细胞Mash1mRNA水平明显下降。结论化学合成siRNA能有效地抑制大鼠骨髓间充质干细胞Mash1的表达。
Objective To screen siRNA sequences that can effectively suppress Mash1 gene expression in rat bone marrow mesenchymal stem cells. Methods According to the design principle of siRNA target sequence, three pairs of siRNAs targeting the Mash1 gene coding region of the rat were designed and synthesized. A pair of unrelated oligonucleotides was used as a negative control. Primary cultured rat bone marrow mesenchymal stem cells and divided into 5 groups: Mash1-1 group, Mash1-2 group, Mash1-3 group, unrelated sequence group, blank group. The expression of Mash1 was detected by RT-PCR at 48h after transfection with cationic liposome LipofectamineTM 2000. Results Mash1 mRNA of rat bone marrow mesenchymal stem cells transfected with sequence 1 and sequence 2 was significantly decreased 48 h after transfection compared with those of sequence 3, unrelated sequence and blank control group. Conclusion Chemically synthesized siRNA can effectively inhibit the expression of Mash1 in rat bone marrow mesenchymal stem cells.