P-20: 3’UTR of Oct-4 Gene that Inserted Downstream of LUC Gene Cause Decrease in Luciferase Count in P19, 5637 and A172 Cell Lines (Pages: 48-49)

Ghiasvand S *, Najafi R , Sadeghizadeh M , Mowla SJ ,


Objective: The 3' UTR is a particular section of Messenger RNA that is not translated but has regulatory role in translation. In many cases gene expression control mechanism occurs at translational stage. In one mechanism protein-mediated interactions between UTRs result in the formation of a RNA loop, Such RNA ‘circularization’ is thought to increase translational efficiency. In another mechanism microRNAs that are a new class of regulatory elements in gene expression bind to complementary sequences in the 3' UTR of multiple target mRNAs, usually resulting in their silencing. Oct-4 as a POU homeobox transcription factor, is preferentially expressed and active in ES cells, and expression appears to be required for maintenance of the undifferentiated state of ES cells. Oct-4 expression has also been detected in adult stem cells as well as a variety of cancer cells such as bladder, liver, stomach and other tumoral cell lines. Oct-4 can be a marker for prognostic diagnosis of many cancers especially for bladder cancer. Materials and Methods: In our study, the Oct-4 3'UTR after isolation and amplification with PCR technique inserted into the pGL3-control reporter vector downstream of LUC gene with cloning method, and then transfected into the P19 (carcinoma stem cell) ,5637 and A172 cell lines with lipofectamine 2000 according to manufacture protocol , then we using Luciferase assay technique to evaluating the regulatory effect of 3'UTR of Oct4 gene in the expression of luciferase gene in under study cell lines. Results: Our data confirmed the regulatory role of Oct-4 gene 3' UTR in under study cell lines. In all cell lines Oct-4 3’UTR cause decrease in luciferase count, especially in P19 cell line we see a significant decrease in luciferase count in contrast with control vector. Conclusion: Regulatory factors such as microRNAs by attaching to this segment of gene can have significant roles in this inhibitory effect. By bioinformatics analysis we predicted 13 MicroRNAs that can attach to mouse Oct-4 3’UTR. Considering the fact that Oct-4 is the most important gene that express in the early embryonic stage and in cancer cell , the regulatory effect of 3'UTR of Oct-4 gene can be a therapeutic target for cancer treatment