siRNA干扰Oct4基因对P19细胞中miR-302启动子活性的影响  

Effect of Oct4 siRNA on expression of miR-302 in P19 cells

作　　者：刘海英[1] 宋巍[2] 石宇[1] 

机构地区：[1]广州医学院第三附属医院生殖医学中心,广东省产科重大疾病重点实验室,广东广州510150 [2]天津市中心妇产科医院

出　　处：《中国优生与遗传杂志》2012年第8期17-18,25,共3页Chinese Journal of Birth Health & Heredity

摘　　要：目的检测siRNA干扰Oct4基因对P19细胞中miR-302启动子活性的影响,探讨维持胚胎癌(EC)细胞及胚胎干细胞(ES)全能性的调控机制。方法采用Western blotting检测Oct4 siRNA对Oct4蛋白的干扰效果;采用双荧光素酶报告基因分析系统检测siRNA干扰Oct4基因对P19细胞中miR-302基因启动子活性的影响。结果对于Oct4蛋白的最适干扰量为2.0μg siRNA1和2.0μg siRNA2;向P19细胞中转染Oct4 siRNA可以降低miR-302启动子的荧光素酶活性。结论 Oct4对miR-302的启动子具有正向调控作用。Objective ： To investigate the effect of small interfering RNA （siRNA） of Oct4 on expression of miR - 302 in P19 cells and elucidate the regulatory mechanism of pluripotency in EC cells and ES cells. Methods ： The interference effect of Oct4 siRNA on the Oct4 protein and the optimal concentration of interference were detected by Western blotting. The role of Oct4 siRNA interference on miR - 302promoter activity was analyzed by dual luciferase reporter assay system. Results ： The optimal concentration of interference of Oct4 siRNA was 2. 0p, g siRNA1 and 2. 0μ g siRNA2. Transfection of Oct4 siRNA to P19 cells decreased the promoter activity of miR -302. Conclusion： The miR -302 cluster promoter was regulated by Oct4 directly.

关 键 词：胚胎癌细胞 Oct4基因 miR-302 RNA干扰 

分 类 号：R730.2[医药卫生—肿瘤]