引用本文:耿艳清,杨德辉,鲁之中,陈雪梅,何俊琳,王应雄,丁裕斌,刘学庆.早孕小鼠胚胎着床窗口期子宫内膜mmu-miR-106b表达规律及其作用[J].重庆医科大学学报,2013,(10):1154~1158
早孕小鼠胚胎着床窗口期子宫内膜mmu-miR-106b表达规律及其作用
Expression rules and potential role of mmu-miR-106b in the endometrium of mouse during embryo implantation
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中文关键词:  胚胎着床  子宫内膜  mmu-miR-106b
英文关键词:embryo implantation  endometrium  mmu-miR-106b
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耿艳清,杨德辉,鲁之中,陈雪梅,何俊琳,王应雄,丁裕斌,刘学庆 重庆医科大学公共卫生与管理学院生殖生物学研究室重庆 400016 
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中文摘要:
      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real-time 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mmu-miR-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。
英文摘要:
      Objective:To explore the expression rules and potential role of mmu-miR-106b in the endometrium mouse during embryo implantation. Methods:Real-time PCR and in situ hybridization were used to detect the expressions of mmu-miR-106b in the en-dometria of pregnant mice on d4,d5 and d6. MTT and flow cytometry were performed to understand its roles in endometrial stromal cell proliferation,cycle and apoptosis after endometrial stromal cells being transfected with mimics and inhibitor. Western blot was car-ried out to predict the potential target genes of mmu-miR-106b by combining anticipated algorithms. Results:Expression of mmu-miR-106b in the endometria on d6 was lower than that on d4(P=0.039) and there was no significant difference in mmu-miR-106b expression between implantation sites and inter-implantation sites of pregnant mice on d5(P=0.606). Mmu-miR-106b was located in endometrial stromal cells. Increased mmu-miR-106b could promote cell proliferation and reduced mmu-miR-106b would increase cell apoptosis. Related target genes of nucleophosmin1,tumor susceptibility gene 101(Tsg101) and phosphatase and tensin homolog deleted on chromosome ten were achieved by miRGen,Targetscan and Pictar screening and expression of Tsg101 was regulated by mmu-miR-106b in endometrial stromal cells of pregnancy mice(P=0.042). Conclusions:Mmu-miR-106b may play an important role in the proliferation of endometrial stromal cells during embryo implantation process by targeting Tsg101.
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