孕激素诱导的microRNA-1a抑制子宫内膜上皮细胞增殖

发布时间：2018-01-09 13:02

本文关键词：孕激素诱导的microRNA-1a抑制子宫内膜上皮细胞增殖　出处：《生理学报》2016年06期 　论文类型：期刊论文

【摘要】：本文旨在研究孕激素诱导micro RNA-1a(mi R-1a)在子宫内膜上皮细胞(endometrial epithelial cells,EECs)的表达及其对EECs增殖的作用。用雌激素(E2组)或雌、孕激素联合(E2P4组)处理去卵巢小鼠,q PCR检测mi R-1a的成熟体mi R-1a-3p的表达;将mi R-1a-3p拟似剂或抑制剂注入各组小鼠的一侧子宫角,另一侧注入拟似剂或抑制剂对照,流式细胞术检测mi R-1a-3p对EECs细胞周期的影响;免疫组化检测mi R-1a-3p对细胞周期蛋白cyclin D2、cyclin E1和cyclin E2表达的影响。在体外实验,将分离培养的原代小鼠EECs分为两组,分别给予雌激素处理(E2组)和雌、孕激素联合处理(E2P4组),q PCR检测mi R-1a-3p的表达;再用mi R-1a-3p拟似剂作用于雌激素预处理细胞,对照组给予非特异性拟似剂,Ed U掺入实验检测细胞增殖活性。体内q PCR结果显示,E2P4组小鼠EECs mi R-1a-3p的表达量相比E2组明显增加(P0.05)。流式细胞术结果显示mi R-1a-3p拟似剂能够将E2处理细胞的细胞周期进程阻滞在G1/S期(P0.05);mi R-1a-3p抑制剂能够部分逆转P4对细胞周期进程的G1/S期阻滞(P0.05)。小鼠子宫组织免疫组化结果显示mi R-1a-3p拟似剂能较明显地抑制cyclin E1和cyclin E2蛋白的表达(P0.05),而对cyclin D2蛋白表达无影响(P0.05)。体外q PCR结果显示E2处理组和E2P4联合处理组细胞中都未检测到mi R-1a-3p表达,体外Ed U掺入实验表明外源性mi R-1a-3p拟似剂能够一定程度抑制雌激素促细胞增殖作用(P0.05)。以上结果表明,孕激素可通过诱导mi R-1a-3p表达引起细胞周期G1/S期阻滞,从而抑制EECs细胞增殖,这种抑制增殖作用与其抑制cyclin E1和cyclin E2蛋白表达有关。
[Abstract]:The aim of this study was to investigate the effects of progesterone-induced micro RNA-1a(mi R-1a on endometrial epithelial cells (. Endometrial epithelial cells. Ovariectomized mice were treated with estrogen estradiol (E2) or estrogen, progesterone combined with E2P4). Q PCR was used to detect the expression of mi R-1a-3p in the mature body of mi R-1a. The miR-1a-3p mimic agent or inhibitor was injected into one side of uterus horn of mice in each group, and the other side was injected with mimic agent or inhibitor control. The effect of mi R-1a-3p on EECs cell cycle was detected by flow cytometry. The effect of miR-1a-3p on the expression of cyclin E1 and cyclin E2 of cyclin cyclin D2 was detected by immunohistochemistry in vitro. The isolated and cultured primary EECs mice were divided into two groups: estradiol treatment group (E2 group), estrogen group and progesterone combined treatment group (E2P4 group). The expression of miR-1a-3p was detected by Q PCR. Then the cells were pretreated with miR-1a-3p mimic agent, and the control group was treated with non-specific mimic agent Ed U incorporation to detect cell proliferation activity. The results of Q PCR in vivo showed that the cells were pretreated with estrogen. The expression of EECs mi R-1a-3p in E2P4 group was significantly higher than that in E2 group (P 0.05). The results of flow cytometry showed that miR-1a-3p mimic agent could block the cell cycle progression of E2 treated cells at G 1 / S phase (P 0.05). Mi R-1a-3p inhibitor can partially reverse the G 1 / S phase arrest of P4 to cell cycle progression (P0.05). The immunohistochemical results of mouse uterus showed that miR-1a-3p mimic agent could significantly inhibit the expression of cyclin E1 and cyclin E2 protein (P0.05). The expression of cyclin D2 protein was not affected by P0.05. The results of Q PCR in vitro showed that neither E2 nor E2P4 could detect the expression of miR-1a-3p in E2P4 cells. In vitro Ed U incorporation test showed that exogenous miR-1a-3p mimic agent could inhibit the effect of estrogen on cell proliferation to a certain extent. Progesterone can inhibit the proliferation of EECs cells by inducing the expression of mi R-1a-3p to block the G _ 1 / S phase of cell cycle. This inhibitory effect is related to the inhibition of cyclin E1 and cyclin E2 protein expression.
【作者单位】：四川大学华西基础医学与法医学院生理学教研室;
【基金】：supported by the National Natural Science Foundation of China(No.31271251)
【分类号】：R714
【正文快照】： 子宫的结构和功能受到雌激素和孕激素的双重调控,雌激素促进子宫内膜上皮细胞(endometrialepithelial cells,EECs)增殖,孕激素可拮抗雌激素对EECs的促增殖效应,并促使其分化为具有分泌功能的组织,以保证胚胎在宫腔内的正常发育和顺利着床。如孕激素作用不足,可能导致内膜增生

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