Expression of Long Non-Coding RNA H19 in the Endometrium of Mice During Peri-Implantation and Its Regulation on Embryo Implantation

IF 2.9 4区 医学 Q1 Medicine
Haixia Li, Ning Su, Ya‐ning Zhu, Wei Wang, Meihong Cai, Sushi Jiang, Xiaohuan Luo, Wei Xia
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Abstract

In this study, we utilized magnetic nanobeads for the extraction of nucleic acids from tissues to investigate the expression levels and correlation between lncRNA H19, miR-612, and their target gene HOXA10 in peri-implantation endometrium of mice. Furthermore, we conducted overexpression or gene knockout experiments on lncRNA H19 to observe its impact on the expression of miR-612 and HOXA10. The targeted binding relationship between lncRNA H19, miR-612, and HOXA10 was detected by dual luciferase reporter assay. The regulatory relationship between lncRNA H19, miR-612, and HOXA10 was verified through silencing or overexpression of these genes. Intrauterine transfection was used to modulate the expression of lncRNA H19 in endometria during pregnancy, followed by the detection of the expression levels of miR-612 and HOXA10 as well as ITGB3 and IGFBP-1 proteins. Compared with non-pregnant mice, we observed a significant upregulation of both lncRNA H19 and HOXA10 in the endometria of pregnant mice, while miR-612 was found to be downregulated (P < 0.05). Further analysis revealed that the expression levels of lncRNA H19 and HOXA10 increased progressively with gestational days, peaking on Day 4 (P < 0.05). Moreover, Through database analysis, we identified binding sites for lncRNA H19-miR-612 as well as HOXA10-miR-612 interactions. The dual-luciferase reporter assay further supported our conjecture that lncRNA H19 could specifically bind the miR-612, which in turn targets HOXA10 to regulate its expression (P < 0.05). In conclusion, regulations of lncRNA H19 and HOXA10 expression contribute to enhancing endometrial receptivity and facilitating decidualization of endometrial stromal cells, ultimately promoting successful embryo implantation.
长非编码 RNA H19 在小鼠子宫内膜着床期的表达及其对胚胎着床的调控
本研究利用纳米磁珠从组织中提取核酸,研究lncRNA H19、miR-612及其靶基因HOXA10在小鼠着床周围子宫内膜中的表达水平及其相关性。此外,我们还对lncRNA H19进行了过表达或基因敲除实验,观察其对miR-612和HOXA10表达的影响。通过双荧光素酶报告实验检测了lncRNA H19、miR-612和HOXA10之间的靶向结合关系。通过沉默或过表达这些基因,验证了lncRNA H19、miR-612和HOXA10之间的调控关系。通过宫腔内转染来调节妊娠期子宫内膜中lncRNA H19的表达,然后检测miR-612和HOXA10以及ITGB3和IGFBP-1蛋白的表达水平。与非妊娠小鼠相比,我们观察到妊娠小鼠子宫内膜中的lncRNA H19和HOXA10均显著上调,而miR-612则下调(P < 0.05)。进一步分析发现,lncRNA H19和HOXA10的表达水平随着妊娠天数的增加而逐渐升高,在第4天达到峰值(P < 0.05)。此外,通过数据库分析,我们确定了lncRNA H19-miR-612以及HOXA10-miR-612相互作用的结合位点。双荧光素酶报告实验进一步支持了我们的猜想,即lncRNA H19可以特异性地与miR-612结合,进而靶向HOXA10以调控其表达(P < 0.05)。总之,lncRNA H19和HOXA10的表达调控有助于提高子宫内膜的容受性和促进子宫内膜基质细胞的蜕膜化,最终促进胚胎成功着床。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
4.30
自引率
17.20%
发文量
145
审稿时长
2.3 months
期刊介绍: Information not localized
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