Androgen-induced alterations in endometrial proteins crucial in recurrent miscarriages.
Androgen-induced alterations in endometrial proteins crucial in recurrent miscarriages.
复制标题
雄激素诱导的子宫内膜蛋白改变对复发性流产至关重要
DOI：
10.18632/oncotarget.24821
复制
发表时间：
2018-05-15
期刊：
影响因子：
--
通讯作者：
Huang HF
中科院分区：
文献类型：
--
作者：
Rahman TU;Ullah K;Guo MX;Pan HT;Liu J;Ren J;Jin LY;Zhou YZ;Cheng Y;Sheng JZ;Huang HF
高雄激素水平损害反复流产妇女子宫内膜容受性。雄激素对子宫内膜的作用机制尚不清楚。我们假设雄激素对复发性流产妇女的子宫内膜有直接影响。在本研究中，我们评估的影响雄激素（A2）在高浓度（10-7 M）的石川细胞相比，生理浓度的雄激素（10-9 M）。为了进行更深入的分析，我们使用全局稳定同位素标记的分析策略，使用iTRAQ试剂，然后进行2D LC-MS/MS。我们确定了175个非冗余蛋白质，并对其中18个进行了定量。差异表达蛋白质（DEPs）分析发现，高雄激素组中有8种蛋白质上调，10种蛋白质下调。这些DEPs通过独创性途径（IPA）分析进行了检查，并确定这些蛋白质可能在复发性流产和子宫内膜容受性中发挥重要作用。此外，蛋白质细胞周期蛋白依赖性激酶抑制剂2a（CDKN 2a），内皮蛋白C受体（EPCR），犰狳重复velocardiofacial（ARVCF）被独立地使用蛋白质印迹法确认。敲低CDKN 2a可显著降低石川细胞中CDKN 2a蛋白的表达水平，并降低细胞的迁移（p < 0.01）、侵袭（p <0.05）、增殖（p <0.05）和对石川细胞单层的Jar球体附着率（p < 0.05）。结果提示，高浓度雄激素可改变子宫内膜发育和胚胎着床相关蛋白的表达水平，这可能是导致子宫内膜容受性受损和流产的原因之一。
High androgen level impairs endometrial receptivity in women experiences the recurrent miscarriage. The mechanism of androgen actions on endometrium is still uncertain. We hypothesized that androgen has a direct effect on the endometrium in women with recurrent miscarriage. In the present study, we assess the impact of androgen (A2) at high concentration (10–7 M) on Ishikawa cells compared with the physiological concentration of androgen (10–9 M). To go into deeper analysis, we use global stable isotopes labeled profiling tactic using iTRAQ reagents, followed by 2D LC-MS/MS. We determine 175 non-redundant proteins, and 18 of these were quantified. The analysis of differentially expressed proteins (DEPs) identified 8 up-regulated proteins and 10 down-regulated in the high androgen group. These DEPs were examined by ingenuity pathway (IPA) analysis and established that these proteins might play vital roles in recurrent miscarriage and endometrium receptivity. In addition, proteins cyclin-dependent kinase inhibitor 2a (CDKN2a), endothelial protein C receptor (EPCR), armadillo repeat for velocardiofacial (ARVCF) were independently confirmed using western blot. Knockdown of CDKN2a significantly decreased the expression level of CDKN2a protein in ishikawa cells, and decreased migration (p < 0.01), invasion (p < 0.05), proliferation (p < 0.05), and the rate of Jar spheroid attachment (p < 0.05) to Ishikawa cell monolayer. The present results suggest that androgen at high concentration could alter the expression levels of proteins related to endometrium development and embryo implantation, which might be a cause of the impaired endometrial receptivity and miscarriage.