Fragile X-related protein 1 (FXR1) regulates cyclooxygenase-2 (COX-2) expression at the maternal-fetal interface
Fragile X-related protein 1 (FXR1) regulates cyclooxygenase-2 (COX-2) expression at the maternal-fetal interface
复制标题
脆性 X 相关蛋白 1 (FXR1) 调节母胎界面的环氧合酶 2 (COX-2) 表达
DOI：
10.1071/rd18037
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发表时间：
2018-10-01
影响因子：
1.9
通讯作者：
Chen, Jin-Hong
中科院分区：
文献类型：
--
作者：
Li, Xiao-Cui;Song, Meng-fan;Chen, Jin-Hong
环氧合酶-2（考克斯-2）在转录后受其mRNA 3 '非翻译区（UTR）富含AU元件（ARE）的调控。然而，迄今为止，考克斯-2在不孕症中的诱导机制尚未完全阐明。本研究旨在探讨滋养层细胞中考克斯-2与脆性X相关蛋白1（FXR 1）的关系。使用定量逆转录聚合酶链反应，我们的研究结果表明，FXR 1 mRNA的表达水平显着降低滋养细胞从复发性流产患者与健康对照组相比，相反，考克斯-2 mRNA的表达水平在患者样本中增加。我们还观察到，FXR 1高表达的人胎盘绒毛在妊娠早期。此外，我们使用蛋白质印迹和免疫荧光法来分析用肿瘤坏死因子α处理的HTR-8细胞中FXR 1和考克斯-2的表达水平;我们观察到，在用FXR 1小干扰RNA处理的HTR-8细胞中，考克斯-2的表达明显增加，而在通过质粒过表达FXR 1的HTR-8细胞中，考克斯-2的表达有效降低。重要的是，生物信息学分析鉴定了考克斯-2的3 '-UTR区域中的FXR 1结合位点，萤火虫荧光素酶报告基因测定分析证实了FXR 1直接结合考克斯-2的3'-UTR区域。ELISA分析表明，过表达FXR 1增强血管内皮生长因子-A和白细胞介素-8在HTR-8细胞的表达，而相反，敲低FXR 1有效地抑制这些影响。总之，本研究结果表明，FXR 1是一种新的考克斯-2调节因子。
Cyclooxygenase-2 (COX-2) is regulated post-transcriptionally by the AU-rich element (ARE) in the 3'-untranslated region (UTR) of its mRNA. However, the mechanism of COX-2 induction in infertility has not been thoroughly elucidated to date. The aim of this study was to examine the association between COX-2 and fragile X-related protein 1 (FXR1) in trophoblasts. Using quantitative reverse transcription polymerase chain reaction, our results showed that FXR1 mRNA expression levels were significantly decreased in trophoblasts from recurrent miscarriage patients compared with healthy controls; conversely, COX-2 mRNA expression levels were increased in patient samples. We also observed that FXR1 was highly expressed in human placental villi during early pregnancy. Furthermore, we used western blotting and immunofluorescence to analyse the expression levels of FXR1 and COX-2 in HTR-8 cells that were treated with tumour necrosis factor cc; we observed that the expression of COX-2 was clearly increased in HTR-8 cells treated with FXR1 small interfering RNA, whereas the expression of COX-2 was effectively decreased in HTR-8 cells with FXR1 overexpressed via a plasmid. Importantly, bioinformatics analysis identified FXR1 binding sites in the 3'-UTR region of COX-2 and firefly luciferase reporter assay analysis verified that FXR1 binds directly to the 3'-UTR region of COX-2. ELISA assays showed that overexpression of FXR1 enhanced vascular endothelial growth factor-A and interleukin-8 expression in HTR-8 cells, whereas conversely, knockdown of FXR1 effectively repressed these effects. In conclusion, the results of this study indicate that FXR1 is a novel COX-2 regulatory factor.