缺氧诱导因子-1α/E1A 结合蛋白 P300 复合物抑制人先兆子痫中缺氧时的 G 蛋白信号转导调节因子 2。

Regulator of G protein signaling 2 is inhibited by hypoxia-inducible factor-1α/E1A binding protein P300 complex upon hypoxia in human preeclampsia.

机构信息

National Clinical Research Center for Child Health of the Children's Hospital, Zhejiang University School of Medicine, Hangzhou 310052, China; Department of Obstetrics, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China.

National Clinical Research Center for Child Health of the Children's Hospital, Zhejiang University School of Medicine, Hangzhou 310052, China.

出版信息

Int J Biochem Cell Biol. 2022 Jun;147:106211. doi: 10.1016/j.biocel.2022.106211. Epub 2022 Apr 14.

DOI:10.1016/j.biocel.2022.106211

PMID:35430356

Abstract

BACKGROUND

Preeclampsia is a pregnancy-related complication that causes maternal and fetal mortality. Despite extensive studies showing the role of hypoxia in preeclampsia progression, the specific mechanism remains unclear. The purpose of this study was to explore the possible mechanism underlying hypoxia in preeclampsia.

METHODS

Human trophoblast-like JEG-3 cell line was used to investigate the molecular mechanisms underlying hypoxia contribution to preeclampsia and the expression correlation of key molecules was examined in human placental tissues. Methods include JEG-3 cell culture and hypoxia induction, RNA isolation and quantitative real-time PCR, transient transfection and dual-luciferase assay, western blot, immunoprecipitation, immunofluorescence staining, cell proliferation assay, chromatin immunoprecipitation assay, obtainment of human placental tissue sample and immunohistochemistry staining.

RESULTS

Hypoxia-Inducible Factor-1α is up-regulated in clinical preeclampsia samples, where Regulator of G Protein Signaling 2 is down-regulated. Mechanistically, Hypoxia-Inducible Factor-1α is induced in response to hypoxia, which up-regulates E1A binding protein P300 expression and thereby forms a Hypoxia-Inducible Factor-1α/E1A binding protein P300 protein-protein complex that binds to the promoter of gene Regulator of G Protein Signaling 2 and subsequently inhibits the transcription of Regulator of G Protein Signaling 2, possibly contributing to the preeclampsia development. In addition, the expression of E1A binding protein P300 is increased in preeclampsia samples, and the expression of Regulator of G Protein Signaling 2 in preeclamptic placentas inversely correlates with the levels of E1A binding protein P300.

CONCLUSION

Our findings may provide novel insights into understanding the molecular pathogenesis of preeclampsia and may be a prognostic biomarker and therapeutic target for preeclampsia.

摘要

背景

子痫前期是一种与妊娠相关的并发症，可导致母婴死亡。尽管大量研究表明缺氧在子痫前期进展中的作用，但具体机制尚不清楚。本研究旨在探讨缺氧在子痫前期中的可能机制。

方法

使用人绒毛膜滋养层样 JEG-3 细胞系研究缺氧对子痫前期的贡献的分子机制，并在人胎盘组织中检查关键分子的表达相关性。方法包括 JEG-3 细胞培养和缺氧诱导、RNA 分离和定量实时 PCR、瞬时转染和双荧光素酶测定、Western blot、免疫沉淀、免疫荧光染色、细胞增殖测定、染色质免疫沉淀测定、获得人胎盘组织样本和免疫组织化学染色。

结果

在临床子痫前期样本中，缺氧诱导因子-1α 上调，而 G 蛋白信号调节因子 2 下调。在机制上，缺氧诱导因子-1α 对缺氧作出反应而上调 E1A 结合蛋白 P300 的表达，从而形成缺氧诱导因子-1α/E1A 结合蛋白 P300 蛋白-蛋白复合物，与 G 蛋白信号调节因子 2 的启动子结合，随后抑制 G 蛋白信号调节因子 2 的转录，可能导致子痫前期的发展。此外，E1A 结合蛋白 P300 在子痫前期样本中的表达增加，而子痫前期胎盘组织中 G 蛋白信号调节因子 2 的表达与 E1A 结合蛋白 P300 的水平呈负相关。