NANOG regulate the JAK/STAT3 pathway to promote trophoblast cell migration and epithelial-mesenchymal transition (EMT) in hypertensive disorders of pregnancy (HDP) through protein interaction with CDK1

IF 2.5 3区医学 Q3 IMMUNOLOGY American Journal of Reproductive Immunology Pub Date : 2024-05-26 DOI:10.1111/aji.13863

Jing Ma, Mingchang Liu, Zhuo Chen, Shiyang Liu, Huijuan Yang, Mengjia Duan

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Abstract

Problem

Hypertensive disorders of pregnancy (HDP) are a common pregnancy disease. NANOG and Cyclin-dependent kinase 1 (CDK1) are essential for regulating the function of cell proliferation and apoptosis. However, the mechanism of action in HDP is yet unclear.

Method

The microarray dataset GSE6573 was downloaded from the GEO database. Emt-related gene set was downloaded from Epithelial-Mesenchymal Transition gene database 2.0 were screened differentially expressed genes by bioinformatics analysis. Pathway Commons and Scansite 4.0 databases were used to predict the interaction between proteins. Placental tissue samples were collected from HDP patients and patients with uneventful pregnancies. RT-qPCR, Western blot and immunohistochemistry were used to detect the expression of NANOG, CDK1, MMP-2, MMP-9, EMT markers and the JAK/STAT3 pathway proteins. Transfection NANOG overexpression/knockdown, and CDK1 knockdown into the human chorionic trophoblast cells (HTR-8/Svneo). CCK-8, Transwell and Wound-healing assay were used to evaluate cell proliferation, invasion and migration. CO-IP and GST pull-down assays were used to confirm the protein interaction.

Results

A total obtained seven EMT-related differentially expressed genes, wherein NANOG, NODAL and LIN28A had protein interaction. In the HDP patients' tissue found that NANOG and CDK1 had lower expression. NANOG overexpression promoted HTR-8/Svneo proliferation, migration and EMT, while NANOG knockdown had the opposite effect. Further a protein interaction between STAT3 and CDK1 with NANOG. NANOG overexpression downregulated the JAK/STAT3 pathway to promote HTR-8/Svneo proliferation, migration and EMT, which was reversed by CDK1 knockdown.

Conclusions

NANOG downregulated the JAK/STAT3 pathway to promote trophoblast cell proliferation, migration and EMT through protein interaction with CDK1.

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NANOG 通过与 CDK1 蛋白相互作用调控 JAK/STAT3 通路，促进妊娠高血压疾病（HDP）中滋养层细胞的迁移和上皮-间质转化（EMT）

问题妊娠高血压疾病（HDP）是一种常见的妊娠疾病。NANOG和细胞周期蛋白依赖性激酶1（CDK1）是调节细胞增殖和凋亡功能的关键。然而，其在 HDP 中的作用机制尚不清楚。方法从 GEO 数据库下载微阵列数据集 GSE6573。从Epithelial-Mesenchymal Transition gene database 2.0中下载与Emt相关的基因集，通过生物信息学分析筛选出差异表达基因。Pathway Commons 和 Scansite 4.0 数据库用于预测蛋白质之间的相互作用。从 HDP 患者和无异常妊娠患者中采集胎盘组织样本。采用 RT-qPCR、Western 印迹和免疫组织化学方法检测 NANOG、CDK1、MMP-2、MMP-9、EMT 标记和 JAK/STAT3 通路蛋白的表达。将 NANOG 过表达/敲除和 CDK1 敲除转染至人绒毛滋养层细胞（HTR-8/Svneo）。CCK-8、Transwell 和伤口愈合试验用于评估细胞的增殖、侵袭和迁移。CO-IP 和 GST pull-down 试验用于确认蛋白质相互作用。结果共发现 7 个与 EMT 相关的差异表达基因，其中 NANOG、NODAL 和 LIN28A 有蛋白相互作用。在HDP患者组织中发现，NANOG和CDK1的表达量较低。NANOG 的过表达促进了 HTR-8/Svneo 的增殖、迁移和 EMT，而 NANOG 的敲除则产生了相反的效果。进一步发现 STAT3 和 CDK1 与 NANOG 之间存在蛋白质相互作用。NANOG 过表达会下调 JAK/STAT3 通路，从而促进 HTR-8/Svneo 的增殖、迁移和 EMT，而 CDK1 的敲除会逆转这种作用。结论 NANOG通过与CDK1蛋白相互作用，下调JAK/STAT3通路，促进滋养层细胞增殖、迁移和EMT。

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来源期刊

American Journal of Reproductive Immunology 医学-免疫学

CiteScore

6.20

自引率

5.60%

发文量

314

审稿时长

2 months

期刊介绍： The American Journal of Reproductive Immunology is an international journal devoted to the presentation of current information in all areas relating to Reproductive Immunology. The journal is directed toward both the basic scientist and the clinician, covering the whole process of reproduction as affected by immunological processes. The journal covers a variety of subspecialty topics, including fertility immunology, pregnancy immunology, immunogenetics, mucosal immunology, immunocontraception, endometriosis, abortion, tumor immunology of the reproductive tract, autoantibodies, infectious disease of the reproductive tract, and technical news.