Department of Genetics, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran.
Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Basic Med Sci. 2014 Jul;17(7):490-6.
OCT4 is a transcription factor required for pluripotency during early embryogenesis and the maintenance of identity of embryonic stem cells and pluripotent cells. Therefore, the effective expression regulation of this gene is highly critical. UTR regions are of great significance to gene regulation. In this study, we aimed to investigate the potential regulatory role played by 5´UTR and 3´UTR of the Oct4 gene in mouse BMSC and P19 cells.
The Oct4 5´UTR and 3´UTR sequences were cloned into pGL3 luciferase plasmid which led to the generation of pGL3 5´-UTR, pGL3 5´&3´-UTRs and pGL3 3´-UTR vectors. The vectors were transfected into BMSC and P19 cells followed by luciferase assay.
The assay of luciferase expression exhibited a direct link between the presence of Oct4 3´- UTR and the decrease of luciferase count in both cell lines; whereas 5´UTR indicated diverse behaviors in two cells. This discrepancy could be explained in view of the difference of cellular contexts in which the Oct4 UTRs act.
This study sheds some light on the role of UTR regions of mouse Oct4 in regulating post-transcriptional gene expression in pluripotent cells. These data represent potential to be used for the development of novel therapeutic approaches for a variety of malignancies.
OCT4 是早期胚胎发生过程中多能性所必需的转录因子，也是胚胎干细胞和多能细胞身份的维持所必需的。因此，该基因的有效表达调控至关重要。UTR 区域对基因调控具有重要意义。在本研究中，我们旨在研究 Oct4 基因的 5'UTR 和 3'UTR 在小鼠 BMSC 和 P19 细胞中可能发挥的调节作用。
将 Oct4 5'UTR 和 3'UTR 序列克隆到 pGL3 荧光素酶质粒中，生成 pGL3 5'UTR、pGL3 5'&3'UTR 和 pGL3 3'UTR 载体。将载体转染到 BMSC 和 P19 细胞中，然后进行荧光素酶测定。
荧光素酶表达测定显示，Oct4 3'UTR 的存在与两种细胞系中荧光素酶计数的减少之间存在直接联系；而 5'UTR 在两种细胞中的表现则不同。这种差异可以从 Oct4 UTR 在不同细胞环境中发挥作用的差异来解释。
本研究揭示了小鼠 Oct4 的 UTR 区域在调节多能细胞中转录后基因表达的作用。这些数据为开发针对多种恶性肿瘤的新型治疗方法提供了潜力。
