人子宫内膜细胞中 miRNAs 的表达及功能分析

摘要： 目的 探索微小 RNAs （miRNAs， miR） -125b、 miR-30b 和 miR-424 在子宫内膜中的表达及功能。方法 收 集自然周期患者（增殖期和分泌期）和促排卵周期患者（高孕组和非高孕组）的子宫内膜标本， 体外分离培养子宫内 膜上皮细胞和间质细胞， 并用免疫荧光验证。实时定量 PCR 检测 miR-125b、 miR-30b 和 miR-424 的表达。结果 增 殖期， 间质细胞中 miR-125b、 miR-30b 和 miR-424 的表达高于上皮细胞； 分泌期， 间质细胞中 miR-125b 和 miR-424 的表达仍高于上皮细胞， 而 miR-30b 的表达低于上皮细胞。上皮细胞中， 分泌期 miR-125b、 miR-30b 和 miR-424 的 表达显著高于增殖期， 而间质细胞中 3 种 miRNAs 的表达差异无统计学意义。HCG 日高孕组上皮细胞中 miR-125b 的表达高于非高孕组， 间质细胞中 miR-30b 的表达高于非高孕组。miR-125b、 miR-30b 和 miR-424 的靶基因功能 分析发现， 其涉及的生物过程主要有细胞迁移、 运动、 细胞间黏附连接等， 主要富集的信号通路包括胰岛素信号通 路、 VEGF 信号通路、 MAPK 信号通路、 焦点黏附和 Wnt 信号通路。结论 miR-125b、 miR-30b 和 miR-424 在子宫内 膜不同细胞类型、 不同时期和 HCG 日高孕酮患者中的表达存在差异， 可能参与子宫内膜容受性的调节。

The expression and function of miRNAs in human endometrial cells

Abstract: Objective To explore the expression and function of miR-125b, miR-30b and miR-424 in endometrial cells. Methods Human endometrial samples were obtained in natural cycles and stimulating cycles. Endometrial epithelial cells (EECs) and endometrial stromal cells (ESCs) were isolated and confirmed by immunofluorescence. The expressions of miR-125b, miR-30b and miR-424 were detected by real-time PCR. Results The expression levels of miR-125b, miR- 30b and miR-424 were higher in proliferative phase in ESCs than those in EECs. And in EECs, the expression levels of miR-125b, miR-30b and miR-424 were significantly up-regulated in secretory phase than in proliferative phase, while it was stable in ESCs. In addition, the expressions of miR-125b in EECs and miR-30b were increased in ESCs in women with elevated progesterone on the day of HCG administration than those of the control. The target genes of miR-125b, miR-30b and miR- 424 mainly participated in cell migration and motion, cell- cell adherens junction and Wnt signaling pathway. Conclusion miR-125b, miR-30b and miR-424 were differently expressed in endometrial cells in different phases, and may participate in regulation of endometrial receptivity.