胚胎癌F9细胞中Oct4在miRNA302启动子上的募集

Oct4 Binds to miRNA302 Cluster Promoter in F9 Mouse Embryonic Carcinoma Cell

摘要: 目的检测胚胎癌F9细胞中Oct4是否能在miR302启动子上募集。方法采用荧光半定量Real-time PCR检测F9细胞中Oct4RNA干扰后Oct4及miR302表达量的变化,采用染色质免疫沉淀技术(CHIP)检测F9 细胞中Oct4在miR302启动子上募集的情况。结果(1)Oct4 RNA干扰后Oct4表达下降了56%,miR302的表达下降了38%,两者具有相似的表达趋势。(2)Oct4能在miR302启动子上募集。结论F9细胞中Oct4及miR302的表达具有相关性,miR302启动子上有Oct4募集,Oct4对miR302启动子具有调节作用,miR302可能是Oct4的靶基因之一。

Abstract: Objective To investigate whether Oct4 binds to the miR302 cluster promoter in the F9 mouse embryonic carcinoma cell. Methods The expression of Oct4 and miR302 in F9 cells after Oct4 siRNA interference were analyzed by Real-time PCR.We examined the binding of Oct4 to the miR302 cluster promoter in F9 cells by ChIP using antibodies against Oct4.The purified DNA was amplified by PCR using miR302 promoter-specific primers. Results After siRNA-mediated knockdown of Oct4,Oct4 levels was decreased 56% and miR302 expression was decreased 38%,we found that Oct4 bound to the putative promoter region of the miR302 cluster. Conclusion These data indicate that Oct4 and miR302 are coordinately expressed in F9 cells,Oct4 binds to the miR302 cluster promoter in the F9 mouse embryonic carcinoma cell.The miR302 may be regulated by Oct4.