Hypoxia Promotes Extravillous Trophoblast Cell Invasion through the Hypoxia-Inducible Factor Urokinase-Type Plasminogen Activator Receptor Pathway
Hypoxia Promotes Extravillous Trophoblast Cell Invasion through the Hypoxia-Inducible Factor Urokinase-Type Plasminogen Activator Receptor Pathway
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DOI：
10.1159/000525851
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发表时间：
2022-01-01
影响因子：
2.1
通讯作者：
Kobayashi，Hiroshi
中科院分区：
文献类型：
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作者：
Shigemitsu，Aiko;Naruse，Katsuhiko;Kobayashi，Hiroshi
目的缺氧是绒毛外滋养层（EVT）细胞的一个常见特征，可在人类胎盘形成的早期阶段促进侵袭。本研究旨在检测缺氧是否会在体外诱导 EVT 细胞的侵袭表型，并探讨潜在的分子机制。设计在缺氧 (5% O 2) 和常氧 (20% O 2) 条件下检查从妊娠前三个月胎盘组织分离的原代 EVT 细胞的侵袭性。方法使用 IncuCyte 通过 Transwell 和伤口愈合侵袭测定来确定侵袭性ZOOM™ 活细胞成像系统。使用蛋白质印迹分析测量缺氧或常氧处理的细胞的尿激酶纤溶酶原激活剂（uPA）、uPA受体（uPAR）和纤溶酶原激活剂抑制剂的蛋白质表达。使用小干扰RNA (siRNA) 评估缺氧诱导因子1 α (HIF-1α) 的敲低。结果缺氧增强EVT 细胞侵袭，但不影响细胞凋亡。缺氧对 EVT 细胞侵袭性的刺激作用与 uPA-uPAR 途径的诱导有关。合成的uPAR抑制剂显着抑制缺氧诱导的EVT细胞侵袭。 siRNA沉默HIF-1α可消除缺氧的刺激作用并抑制uPAR表达的上调，表明HIF-1α-uPAR信号是缺氧诱导EVT细胞侵袭的关键介质。需要进行进一步的实验来阐明HIF-1α-uPAR信号通路。结论EVT侵袭的低氧调节早期事件可能是由HIF-1α-uPAR通路介导的。
ObjectivesHypoxia is a common feature of extravillous trophoblast (EVT) cells that promote invasion during the early stages of human placentation. This study aimed to examine whether hypoxia-induced an invasive phenotype in EVT cells in vitro and explore the underlying molecular mechanisms.DesignThe invasiveness of primary EVT cells isolated from the first trimester placental tissues during weeks 5–8 of gestation was examined under hypoxic (5% O 2) and normoxic (20% O 2) conditions.MethodsInvasiveness was determined by transwell and wound-healing invasion assays using the IncuCyte ZOOM™ Live-Cell Imaging System. Protein expression of the urokinase plasminogen activator (uPA), uPA receptor (uPAR), and plasminogen activator inhibitor of hypoxia or normoxia-treated cells was measured using Western blot analysis. Knockdown of hypoxia-inducible factor-1 alpha (HIF-1α) was assessed using small interfering RNA (siRNA).ResultsHypoxia enhanced EVT cell invasion but did not affect apoptosis. The stimulatory effect of hypoxia on EVT cell invasiveness was associated with induction of the uPA-uPAR pathway. The synthetic inhibitor of uPAR significantly inhibited hypoxia-induced EVT cell invasion. Silencing of HIF-1α by siRNA abolished the stimulatory effect of hypoxia and inhibited the upregulation of uPAR expression, suggesting that the HIF-1α-uPAR signal is the key mediator for hypoxia-induced EVT cell invasion. Further experiments need to be performed to elucidate the HIF-1α-uPAR signal pathways.ConclusionsThe low oxygen-regulated early events of EVT invasion may be mediated by the HIF-1α-uPAR pathway.