化学合成siRNA对大鼠骨髓间充质干细胞Mash1基因的抑制效应  

The inhibitive effect of chemically synthetic siRNA on the expression of Mash1 of bone marrow stromal stem cells in vitro

作　　者：贾晨光[1,2] 刘卫平[1] 龙乾发[1,3] 刘远杨[1] 伊西才[1] 汪凯[1] 王剑博[1] 阳昊[1] 

机构地区：[1]第四军医大学附属西京医院神经外科,西安710032 [2]湖南省株洲市中心医院 [3]陕西省西安市中心医院神经外科

出　　处：《立体定向和功能性神经外科杂志》2012年第5期265-269,共5页Chinese Journal of Stereotactic and Functional Neurosurgery

基　　金：全军医药卫生科研基金资助项目(编号:10MA026)

摘　　要：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 select the effective sequence of siRNA in inhibiting the expressing of Mash1 of bone marrow stromal stem cells. Methods According to the principle of target sequence of siRNA,we designed three homologous sequences of the complete cds of rattus norvegicus Mash1 mRNA and synthesized siRNA chemically in vitro.A control siRNA which is independent of the genome of rat was also designed and chemically synthesized.Primary bone marrow stromal stem cells of rat were cultured and divided in to 5 groups： Mash1-1 group,Mash1-2group, Mash1-3 group,control siRNA group and untransfected group.Transfection was carried out using LipofectamineTM 2000,a kind of cationic liposome.RT-PCR was employed to detect the change of the expression of Mash1 48h after transfection. Results Compared with the cells that were transfected with siRNA of sequence 3 or nonsense oligonucleotide and the cells that were not transfected,the cells were transfected with siRNA of sequence 1 or 2 shows a lower level of Mash1 mRNA detected by RT-PCR. Conclusion Chemically synthetic siRNA can be used efficiently to inhibit the expression of Mash1 in bone marrow stromal stem cells.

关 键 词：骨髓间充质干细胞 Mash1 SIRNA 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]