Activating transcription factor 3 promotes embryo attachment via up-regulation of leukemia inhibitory factor in vitro.
Activating transcription factor 3 promotes embryo attachment via up-regulation of leukemia inhibitory factor in vitro.
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体外激活转录因子 3 通过上调白血病抑制因子促进胚胎附着
DOI：
10.1186/s12958-017-0260-7
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发表时间：
2017-06-02
期刊：
影响因子：
--
通讯作者：
Sun H
中科院分区：
文献类型：
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作者：
Cheng X;Liu J;Shan H;Sun L;Huang C;Yan Q;Jiang R;Ding L;Jiang Y;Zhou J;Yan G;Sun H
背景接受性子宫内膜是着床过程中母胚分子通讯的基础。然而，子宫内膜容量的具体分子调控机制仍然知之甚少。在这里，我们研究了激活转录因子3（ATF 3）在人类胚胎attachment in vitro.MethodsImmunohistochemistry（IHC）中的表达和ATF 3的功能效应，用于评估人类胚胎ATF 3的表达模式。应用实时荧光定量PCR（qRT-PCR）、免疫印迹和免疫荧光（IF）研究雌激素（E2）和醋酸甲羟孕酮（MPA）处理后石川细胞中ATF 3的表达。通过qRT-PCR和western blotting检测ATF 3增强或抑制后LIF（白血病抑制因子）的表达，并通过荧光素酶报告基因分析和ChIP-PCR证实ATF 3对LIF的调控机制。子宫内膜上皮细胞的能力通过BeWo球状体与石川细胞的体外粘附模型进行评估。结果ATF 3定位于人子宫内膜上皮细胞和间质细胞，在E2和MPA的诱导下，ATF 3在石川细胞中的表达明显增强。腺病毒介导的ATF 3在石川细胞中的过表达激活LIF启动子活性并增强其表达。因此，通过抗体阻断试验，用针对LIF的特异性抗体预处理抑制由ATF 3促进的BeWo球体粘附的刺激。结论ATF 3可能通过上调白血病抑制因子而对人子宫内膜容受性和体外胚胎附着起重要作用。号 2013-081-01 . 2013年12月10日注册。
BackgroundA receptive endometrium is essential for maternal-embryonic molecular communication during implantation. However, the specific molecular regulatory mechanisms of the endometrial capacity remain poorly understood. Here, we examined activating transcription factor 3 (ATF3) expression in human endometria and the functional effect of ATF3 on embryo attachment in vitro.MethodsImmunohistochemistry (IHC) was used to assess the ATF3 expression patterns in human endometria. Quantitative real-time PCR (qRT-PCR), western blotting and immunofluorescence (IF) studies were applied to explore ATF3 expression in Ishikawa cells upon estrogen (E2) and medroxyprogesterone acetate (MPA) treatment. qRT-PCR and western blotting were performed to inspect LIF (leukemia inhibitory factor) expression after enhancement or inhibition of ATF3, and a luciferase reporter assay and ChIP-PCR were used to confirm the regulatory mechanism of ATF3 to LIF. Endometrial epithelial capacity was assessed by an in vitro model of attachment of BeWo spheroids to Ishikawa cells. Western blotting was performed to compare the expression of ATF3 in endometrial samples of recurrent implantation failure (RIF) patients with that in samples from fertile women (FER) who had undergone no less than one successful embryo transplantation.ResultsATF3 was located in human endometrial epithelial cells and stromal cells and was significantly induced by E2and MPA in Ishikawa cells. Adenovirus-mediated overexpression of ATF3 in Ishikawa cells activated LIF promoter activity and enhanced its expression. Accordingly, the stimulation of BeWo spheroid adhesion promoted by ATF3 was inhibited by pretreatment with a specific antibody against LIF via the antibody-blocking assay. Moreover, ATF3 was aberrantly decreased in the endometria of RIF patients.ConclusionsOur findings suggest that ATF3 plays a significant role in regulating human endometrial receptivity and embryo attachment in vitro via up-regulation of leukemia inhibitory factor.Trial registrationConstruction and management of the Nanjing multi-center biobank. No. 2013-081-01 . Registered 10 Dec. 2013.