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目的:探讨mmu-miR-106b在早孕小鼠胚胎着床过程中的表达规律,阐明其对胚胎着床的调控作用。方法:应用realtime PCR和原位杂交检测mmu-miR-106b在早孕小鼠孕4 d、孕5 d及孕6 d子宫内膜中的表达;子宫内膜基质细胞转染mmumiR-106b的模拟物和抑制剂后,MTT和流式细胞仪检测细胞的增殖凋亡情况;结合靶基因预测数据库,利用Western blot确定mmu-miR-106b在子宫内膜中的靶基因。结果:mmu-miR-106b在小鼠孕6 d子宫内膜的表达较孕4 d明显下调(P=0.039),孕5 d着床点与着床旁组织表达无明显差异(P=0.606),定位于子宫内膜基质细胞;上调mmu-miR-106b会促进子宫内膜基质细胞的增殖,下调其表达会促进细胞凋亡;通过miRGen、Targetscan、Pictar数据库筛查获得与胚胎着床相关的靶基因核磷蛋白1、肿瘤易感基因101(tumor susceptibility gene 101,Tsg101)和10号染色体缺失的同源性磷酸酶张力蛋白等,其中Tsg101的表达受到mmu-miR-106b负调控(P=0.042)。结论:mmu-miR-106b可能通过靶向Tsg101,影响胚胎着床过程中子宫内膜基质细胞增殖,对胚胎着床发挥调控作用。
OBJECTIVE: To investigate the expression of mmu-miR-106b during embryo implantation in early pregnancy and to clarify its regulation on embryo implantation. METHODS: The expression of mmu-miR-106b was detected in real-time PCR and in situ hybridization in the endometrium of pregnant mice on the 4th, 5th and 6th day of pregnancy. The endometrial stromal cells were transfected into the mimics of mmumiR-106b And inhibitor, the proliferation and apoptosis of cells were detected by MTT and flow cytometry. Combined with target gene prediction database, the target gene of mmu-miR-106b in endometrium was determined by Western blot. Results: The expression of mmu-miR-106b in the endometrium of mice on the 6th day of pregnancy was significantly lower than that on the 4th day of pregnancy (P = 0.039). There was no significant difference in the expression of the mmu-miR- , Which is located in endometrial stromal cells; upregulation of mmu-miR-106b can promote the proliferation of endometrial stromal cells, and downregulation of its expression can promote cell apoptosis; screening with miRGen, Targetscan, Pictar database and embryo implantation related Targeted nucleophosmin 1, tumor susceptibility gene 101 (Tsg101) and homologous phosphatase tensility protein deleted on chromosome 10, among which the expression of Tsg101 was negatively regulated by mmu-miR-106b (P = 0.042). Conclusion: The effect of mmu-miR-106b on the endometrial stromal cell proliferation may be mediated by targeting Tsg101, which may play an important role in embryo implantation.