NANOG 通过与 CDK1 的蛋白相互作用调节 JAK/STAT3 通路，促进妊娠期高血压疾病（HDP）中滋养细胞的迁移和上皮间质转化（EMT）。
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.
机构信息
Department of Reproduction and Genetics, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.
Yunnan Maternal and Child Health Care Hospital, Kunming, Yunnan, China.
出版信息
Am J Reprod Immunol. 2024 May;91(5):e13863. doi: 10.1111/aji.13863.
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.
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）对于调节细胞增殖和凋亡功能是必不可少的。然而，其在 HDP 中的作用机制尚不清楚。
方法
从 GEO 数据库中下载微阵列数据集 GSE6573。通过生物信息学分析筛选差异表达基因，从 Epithelial-Mesenchymal Transition gene database 2.0 下载 EMT 相关基因集。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 患者和无并发症妊娠患者中收集胎盘组织样本。使用 RT-qPCR、Western blot 和免疫组织化学检测 NANOG、CDK1、MMP-2、MMP-9、EMT 标志物和 JAK/STAT3 通路蛋白的表达。转染 NANOG 过表达/敲低和 CDK1 敲低到人绒毛滋养细胞（HTR-8/Svneo）中。使用 CCK-8、Transwell 和划痕愈合实验评估细胞增殖、侵袭和迁移。使用 CO-IP 和 GST 下拉实验来确认蛋白质相互作用。
结果
总共获得了七个 EMT 相关的差异表达基因，其中 NANOG、NODAL 和 LIN28A 具有蛋白质相互作用。在 HDP 患者的组织中发现 NANOG 和 CDK1 的表达较低。NANOG 过表达促进 HTR-8/Svneo 增殖、迁移和 EMT，而 NANOG 敲低则有相反的效果。进一步证实了 STAT3 和 CDK1 与 NANOG 的蛋白相互作用。NANOG 过表达下调 JAK/STAT3 通路，促进 HTR-8/Svneo 增殖、迁移和 EMT，CDK1 敲低可逆转这一作用。
结论
NANOG 通过与 CDK1 的蛋白相互作用下调 JAK/STAT3 通路，促进滋养细胞增殖、迁移和 EMT。
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