| MiR-182 regulates growth and development in animals.HOXA10 plays an important regulatory role in embryo implantation.HOXA10 was a target gene of mi R-182 as predicted by Bioinformatics analysis.Therefore,studies on the regulation of miR-182 on HOXA10 in endometrial epithelial cells(EEC)will reveal the molecular mechanism of miR-182 during embryo implantation in dairy goats.The present study was performed in Guanzhong dairy goat.Firstly,the expressions of mi R-182 and HOXA10 were detected between receptive and non-receptive endometria.Secondly,the effect of miR-182 on HOXA10 was analyzed,and the regulation of miR-182 on EEC was evaluated.The main results were showed as follows:1.RT-qPCR was used to detect the expression patterns of miR-182 and HOXA10 between receptive and non-receptive endometria.The results showed that the miR-182 and HOXA10 were highly expressed during the receptive phase of goat endometrium.2.Dual-luciferase reporter vector assay,RT-qPCR and western blot were used to verify the regulation of miR-182 on HOXA10.MiR-182 mimics and dual-luciferase reporter vectors were co-transfected into 293 T cells,and luciferase activity was detected.The results showed that the miR-182 mimic group had significantly lower luciferase activity than that of the NC group(P <0.01),whereas miR-182 mimic group didn’t alter luciferase activity than that of the mutant group.These results suggested that miR-182 regulated HOXA10 by combining HOXA10 3′ UTR.The results of RT-qPCR and western blot showed that miR-182 inhibited the HOXA10 expression at mRNA and protein levels.3.The effect of miR-182 on EEC was detected by western blot,cell cycle and cell apoptosis.The results showed that mi R-182 had no effect on cell cycle of EEC but induced EEC apoptosis.Further studies showed miR-182 up-regulated the expression of Caspase-3 and Fas,and down-regulated the expression of BCL-2.MiR-182 could inhibit the mRNA and protein expression of HOXA10,and induce EEC apoptosis. |
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