Abstract: Objective To verify the targeted-regulating effect of hsa-miR-1291 on ARHGAP29 gene. Methods PCR was used to amplify the 3'UTR sequence of the target gene ARHGAP29, using the genome DNA of 293T cells as template and the amplification primer designed based on the 3'UTR sequence information of the gene. After cloning the amplification of ARHGAP29-WT 3'UTR or ARHGAP29-MUT 3'UTR into the pmiR-RB-Report vector, dual-luciferase reporter system was conducted. For further detection of relative fluorescence values, hsa-miR-1291 mimic or Non-target Control were respectively co-transfected into 293T cells with ARHGAP29-WT 3'UTR pmiR-RB-Report(wild-type vector) or ARHGAP29-MUT 3'UTR pmiR-RB-Report(mutant vector). Results ARHGAP29-WT 3'UTR pmiR-RB-Report vector (wild-type vector) and ARHGAP29-MUT 3'UTR pmiR-RB-Report vector (mutant vector) were successfully constructed. Fluorescence test showed that after transfection with wild-type vector, fluorescence expression decreased significantly in hsa-miR-1291 mimic group compared with non-target control (NC) group (0.67±0.04 vs 1.00±0.08, P=0.014). After transfection with hsa-miR-1291 mimic, the fluorescence expression of the mutant vector increased compared with that of the wild-type vector (1.20±0.05 vs 0.67±0.04, P<0.001). Conclusion These data suggested that has-miR-1291 may have a targeted regulatory effect on ARHGAP29.

Key words: dual luciferase reporter system, hsa-miR-1291, ARHGAP29, intrauterine adhesions