Abstract:

OBJECTIVE: To investigate whether Oct4 could activate miR-302 promoter in the F9 mouse embryonic carcinoma cells. METHODS：PGL3-P-miR-302 was created by cloning the PCR- amplified region of the miR-302 putative promoter.  PGL3-P-miR-302 promoter reporter plasmids (0.5  μg) were co-transfected with 0.5 μg or 1.0 μg Oct4 expression vector into 293T cells. PGL3-P-miR-302 promoter reporter plasmids (0.5 μg) were co-transfected with Oct4 siRNA(2.0 μg siRNA1 and 2.0 μg siRNA2) into F9 cells. Luciferase activity was assayed. RESULTS：PGL3-P-miR-302 was created，and the cell-specific activity of this promoter was identified by its  transfection into F9 cells. With 0.5 μg  and 1.0 μg Oct4 overexpression in 293T cells，miR-302 promoter activity increased 2.6-fold and 4.3-fold，respectively. After siRNA-mediated knockdown of Oct4，miR-302 luciferase activity fell to 68% compared with control siRNA. CONCLUSION：This study showed that the miR-302 cluster acted downstream of the Oct4 regulation network in F9 cells.

Key words: Oct4, miR-302 cluster, embryonic carcinoma cells, promoter