CDX1 restricts the invasion of HTR-8/SVneo trophoblast cells by inhibiting MMP-9 expression
CDX1 restricts the invasion of HTR-8/SVneo trophoblast cells by inhibiting MMP-9 expression
复制标题
CDX1通过抑制MMP-9的表达来限制HTR-8/SVneo滋养层细胞的侵袭。
DOI：
10.1016/j.placenta.2014.04.011
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发表时间：
2014-07-01
期刊：
影响因子：
3.8
通讯作者：
Wang, X.
中科院分区：
文献类型：
--
作者：
Jia, R. -Z.;Rui, C.;Wang, X.
导读：早发性先兆子痫（PE）的发病机制通常被认为是滋养细胞侵袭子宫肌动脉受损，导致胎盘功能不全。最近，我们报道了CDX1在人子痫前期胎盘中高甲基化。然而，CDX1是否参与滋养细胞侵袭尚不清楚。方法：通过稳定转染CDX1，研究CDX1在HTR-8/SVneo外滋养细胞中的功能。使用CDX1稳定转染细胞系，我们使用QCM ECMatrix 24孔试剂盒检测细胞侵袭。采用MIT（344,5 -二甲基噻唑-2-y1]- 2,5 -二苯基溴化四唑）测定法检测细胞活力。采用定量RT-PCR和western blotting分析下游靶基因和蛋白的表达变化。为了破坏PI3K/AKT信号，我们使用了PI3K抑制剂perifosine。结果：细胞侵袭实验表明CDX1限制滋养细胞的侵袭。相反，累积细胞数量的定量显示CDX1不影响细胞增殖。Western blotting分析和实时荧光定量PCR显示，过表达cdx1的细胞中MMP-9表达降低，而TIMP-1表达升高。然而，过表达CDX1并不影响HTR-8/SVneo滋养细胞中的PI3K/AKT信号传导。相反，CDX1受PI3K/AKT信号通路调控。结论：总之，我们发现在滋养细胞中，CDX1独立于PI3K/AKT信号通路减少侵袭。此外，CDX1与PI3K/AKT信号一起调节滋养细胞侵袭。我们得出结论，CDX1通过独立于PI3K/AKT通路抑制MMP-9的表达来限制HTR-8/SVneo滋养细胞的侵袭。(C) 2014 Elsevier Ltd.版权所有。
Introduction: Pathogenesis of early-onset preeclampsia (PE) is generally recognized by impaired trophoblast invasion of the myometrial arteries, which results in placental insufficiency. Recently, we reported that CDX1 is hypermethylated in the human preeclampsia placenta. However, whether CDX1 participates in trophoblast invasion has not been clearly elucidated.Methods: We investigated the function of CDX1 in the extravillous trophoblast cell line HTR-8/SVneo using stable transfection of CDX1. Using a CDX1 stable transfected cell line, we determined the cell invasion using a QCM ECMatrix 24-well kit. The cell viability was detected using an MIT (344, 5-dimethylthiazol-2-y1]-2, 5-diphenyltetrazolium bromide) assay. Quantitative RT-PCR and western blotting analyses were performed to examine the changes in the expression of downstream target genes and proteins. To disrupt PI3K/AKT signaling, we used the PI3K inhibitor perifosine.Results: Cell invasion assays demonstrated that CDX1 restricts trophoblast cell invasiveness. In contrast, quantification of cumulative cell numbers revealed that CDX1 did not affect cell proliferation. Western blotting analysis and quantitative real time PCR demonstrated that MMP-9 expression was reduced, whereas TIMP-1 expression was increased in CDX1-overexpressed cells. However, overexpression of CDX1 did not affect PI3K/AKT signaling in HTR-8/SVneo trophoblast cells. In contrast, CDX1 was regulated by the PI3K/AKT signaling pathway.Conclusions: Altogether, we found that in trophoblast cells, CDX1 reduced invasion independently of the PI3K/AKT signaling pathway. Furthermore, CDX1 functions in concert with PI3K/AKT signaling to regulate trophoblast invasion. We concluded that CDX1 restricts the invasion of HTR-8/SVneo trophoblast cells by inhibiting MMP-9 expression independently of the PI3K/AKT pathway. (C) 2014 Elsevier Ltd. All rights reserved.