Expression Of Mmu-miRNA-100 In The Endometrium Of Early Pregnant Mouse During Embryo Implantation

Embryo implantation is a complex progress,which is a multifactor-related process.The endometrium decidualization is an important feature of normal implantation.Some factors has regulated this process,and micro RNAs(miRNAs)play significant function in the accurate regulation of differential expression of genes.According our team previously study have demonstrated the expression of mmu-miR-100 on day 5 in endometrium of early pregnant mouse with miRNAs chip technology and there was significant difference in mmu-miR-100 expression between inter-implantation sites and implantation sites.Objective to study on mmu-miR-100,we tend to explore its regulation in endometrium of mice in embryo implantation which in early pregnant stage.It will bring the new thinking of diagnosis and treatment infertility.Methods1.we used Real-time fluorescence quantitative polymerase chain reaction to find the microarray results,and we apply in situ hybridization to reveal the location of mmu-miR-100 in endometrium of early pregnant mice that on day 1,day4,day5 and day 6 during embryo implantation.2.Endometrial stromal cells of early pregnant mice were applied to culture,mimics and inhibitor of mmu-miR-100 were used to transiently transfect into stromal cells.3.we have performed flow cytometry to understand the effect of mmu-miR-100 on endometrial stromal cells growth cycle and apoptosis of early pregnant mice.4.In the study,target genes prediction database of miRNAs was applied to try to find mmu-miR-100 potential target genes.Results1.FQ-PCR results demonstrated that the expression of mmu-miR-100 in inter-implantation sites and is clearly higher compared with in implantation sites(P<0.05),which consistent with the previous microarray analysis.2.Results of in situ hybridization verified that the mmu-miR-100 was located in the stromal cells on day 5 group,and there was almost no expression in the luminal epithelium and glandular epithelium.In day 6 group,expression intensity of pregnant mice of mmu-miR-100 were detected in the stromal cells and which located in secondary decidual zone mostly.3.Stromal cells were cultured and transfected mimics,flow cytometry results showed that the apoptosis rate of stromal cells increasedsignificantly.However,there are no changes in cell proliferation.4.Combining bioinformatics analysis and previous experimental results,predicted target genes of mmu-miR-100 are suggested that IGF2,PLK1,mTOR,FKBP51,FRAP1 et al.Conclusion1.There was significant difference in mmu-miR-100 expression between in implantation sites and inter-implantation sites on day 5 group and day 6 group.Therefore,the expression of implantation sites of mmu-miR-100 was in endometrial decidual zone.2.Mmu-miR-100 may effect endometrial stromal cells apoptosis and then regulate stromal cells decidualization.