KLF6 negatively regulates HIF-1α in extravillous trophoblasts under hypoxia

IF 3 2区 医学 Q2 DEVELOPMENTAL BIOLOGY Placenta Pub Date : 2024-09-05 DOI:10.1016/j.placenta.2024.09.002
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Abstract

Introduction

HIF-1α, the master regulator of hypoxia cellular response, is stabilized under low oxygen levels and degraded in the presence of oxygen but its transcription, translation, and degradation are tightly regulated by numerous pathways. KLF6 is a transcription factor involved in proliferation, differentiation, and apoptosis in several cell systems. Under hypoxia it is upregulated in a HIF-1α-dependent manner in extravillous trophoblasts. Considering the importance of hypoxia modulation of EVT behavior through HIF1-α we aimed to study whether KLF6 modulates HIF-1α expression in HTR8/SVneo cells.

Methods

HTR8/SVneo cells were cultured in a 1 % oxygen chamber or in 3D format where a spontaneous oxygen gradient is generated. qRT-PCR and Western blot were performed to analyze mRNA and protein expression, respectively. SiRNA, shRNA, or plasmids were used to down- or up-regulate gene expression. Wound healing assay was performed under hypoxia to evaluate migration. The NFκB pathway was modulated with dominant negative mutants and a chemical inhibitor. Cobalt chloride was used to block HIF-1α degradation.

Results

KLF6 up- and down-regulation in HTR8/SVneo cells exposed to acute hypoxia revealed a negative regulation on HIF-1α. KLF6 silencing led to a partially HIF-1α-dependent increase in MMP9 and VEGF. The NF-κB pathway and HIF-1α degradation were involved in KLF6-dependent HIF-1α regulation. HTR8/SVneo-3D culture showed that KLF6 negatively regulates HIF-1α in a microenvironment with naturally generated hypoxia.

Discussion

Present results reveal that KLF6 contributes to a fine tune modulation of HIF-1α level under hypoxia. Thus, sustaining a HIF-1α homeostatic level, KLF6 might contribute to control EVT adaptation to hypoxia.

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KLF6 在缺氧条件下负向调节体外滋养细胞中的 HIF-1α
导言HIF-1α是缺氧细胞反应的主调节因子,在低氧水平下稳定,在有氧条件下降解,但其转录、翻译和降解受到多种途径的严格调控。KLF6 是一种转录因子,参与多个细胞系统的增殖、分化和凋亡。在缺氧条件下,它在体外滋养细胞中以 HIF-1α 依赖性方式上调。考虑到缺氧通过 HIF1-α 调节 EVT 行为的重要性,我们旨在研究 KLF6 是否会调节 HTR8/SVneo 细胞中 HIF-1α 的表达。使用 SiRNA、shRNA 或质粒来下调或上调基因表达。在缺氧条件下进行伤口愈合试验,以评估迁移情况。使用显性负突变体和化学抑制剂调节 NFκB 通路。结果 KLF6在暴露于急性缺氧的HTR8/SVneo细胞中的上调和下调显示了对HIF-1α的负调控。KLF6沉默导致MMP9和血管内皮生长因子的增加部分依赖于HIF-1α。NF-κB途径和HIF-1α降解参与了KLF6依赖性HIF-1α调控。HTR8/SVneo-3D培养显示,在自然产生的缺氧微环境中,KLF6负向调节HIF-1α。因此,维持 HIF-1α 的平衡水平,KLF6 可能有助于控制 EVT 对缺氧的适应。
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来源期刊
Placenta
Placenta 医学-发育生物学
CiteScore
6.30
自引率
10.50%
发文量
391
审稿时长
78 days
期刊介绍: Placenta publishes high-quality original articles and invited topical reviews on all aspects of human and animal placentation, and the interactions between the mother, the placenta and fetal development. Topics covered include evolution, development, genetics and epigenetics, stem cells, metabolism, transport, immunology, pathology, pharmacology, cell and molecular biology, and developmental programming. The Editors welcome studies on implantation and the endometrium, comparative placentation, the uterine and umbilical circulations, the relationship between fetal and placental development, clinical aspects of altered placental development or function, the placental membranes, the influence of paternal factors on placental development or function, and the assessment of biomarkers of placental disorders.
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