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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(P<0.05)。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(P<0.05);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(P<0.05)。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(P<0.05),而对cyclin D2蛋白表达无影响(P>0.05)。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(P<0.05)。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。
This article aims to investigate the expression of miR-1a in progesterone-induced endometrial epithelial cells (EECs) and its effect on the proliferation of EECs. Ovariectomized mice were treated with estrogen (group E2) or estrogen and progesterone (group E2P4), and mi R-1a-3p Like agent or inhibitor was injected into the uterine horn on one side of the mice and the pseudo-agent or inhibitor control on the other side. The effect of mi R-1a-3p on the cell cycle of EECs was detected by flow cytometry. Effect of mi R-1a-3p on cyclin D2, cyclin E1 and cyclin E2 expression. In vitro experiments, the primary cultured mouse EECs were divided into two groups and treated with estrogen (group E2) and estrogen and progesterone (group E2P4) respectively. The expression of mi R-1a-3p was detected by q-PCR The mi R-1a-3p mimics were used to pre-treated estrogen cells. The control group was given nonspecific mimics. Ed U incorporation assay was used to test the cell proliferative activity. In vivo qPCR results showed that the expression of mi R-1a-3p in EECs of E2P4 group was significantly increased compared with E2 group (P <0.05). Flow cytometry showed that mi R-1a-3p mimics blocks the cell cycle progression of E2-treated cells in G1 / S phase (P <0.05); mi R-1a-3p inhibitor partially reversed P4 G1 / S arrest of cell cycle progression (P <0.05). The results of immunohistochemistry in mouse uterus showed that mi R-1a-3p mimics could significantly inhibit the expressions of cyclin E1 and cyclin E2 (P <0.05), but had no effect on the expression of cyclin D2 (P> 0.05) . In vitro q PCR results showed that mi R-1a-3p expression was not detected in the cells treated with E2 and E2P4, and in vitro Ed U incorporation assay showed that exogenous mi R-1a-3p mimics inhibited to a certain extent Estrogen promoted cell proliferation (P <0.05). The above results show that progesterone can inhibit the proliferation of EECs by inhibiting the expression of mi R-1a-3p and arresting the G1 / S phase of cell cycle, which is related to the inhibition of cyclin E1 and cyclin E2 protein expression.