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.
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
