DNA methylation-associated repression of MEST/PEG1 expression contributes to the invasion of extravillous trophoblast cells
DNA methylation-associated repression of MEST/PEG1 expression contributes to the invasion of extravillous trophoblast cells
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DNA 甲基化相关的 MEST/PEG1 表达抑制有助于绒毛外滋养层细胞的侵袭。
DOI：
10.1016/j.placenta.2016.08.093
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发表时间：
2016-10-01
期刊：
影响因子：
3.8
通讯作者：
Qi, Hong-Bo
中科院分区：
文献类型：
--
作者：
Peng, Wei;Chen, Ying;Qi, Hong-Bo
简介：绒毛外细胞滋养层细胞（EVT）侵入母体子宫蜕膜和血管系统对人类胎盘发育和妊娠维持至关重要。印迹基因MEST/PEG 1与滋养层细胞的发育有关，但MEST在EVT侵袭及伴随的早孕并发症中的作用尚不完全清楚。方法：采用免疫荧光、免疫组织化学和Western blot方法检测MEST蛋白的表达和定位。应用靶向两种MEST亚型的特异性小干扰RNA（siRNA）来沉默绒毛外外植体和HTR 8/SVneo细胞中的MEST表达。使用Matrigel侵袭、Transwell迁移测定和xCELLigence系统评估细胞侵袭和迁移。采用亚硫酸氢盐测序聚合酶链反应（BSP）检测MEST基因启动子DNA甲基化水平。结果：MEST蛋白在早孕胎盘EVT及侵袭性EVT细胞系HTR-8/Svneo和HPT-8中均高表达。在细胞滋养层细胞（CTB）和绒毛膜癌衍生的CTB细胞系JEG-3中发现弱MEST表达。特异性siRNA敲低MEST表达显著降低HTR-8/Svneo细胞的侵袭和迁移以及绒毛外外植体的生长，这与Twist、N-cadherin和Vimentin的下调有关。结论：MEST蛋白表达抑制与MEST亚型2启动子甲基化有关，MEST亚型2启动子甲基化可能是导致稽留流产的病理机制之一。本研究结果为稽留流产的发生提供了可能的病理机制。(C)2016爱思唯尔有限公司版权所有
Introduction: The invasion of extravillous cytotrophoblasts (EVTs) into the maternal uterine decidua and vasculature is critical for human placenta development and pregnancy maintenance. The imprinted gene MEST/PEG1 has been implicated in trophoblast development; however, the role of MEST in EVT invasion and the accompanying early pregnancy complications are not fully understood.Methods: Western blot, immunofluorescence and immunohistochemistry were used to detect MEST protein expression and localization by using antibodies recognize 2 reported isoforms. Specific small interference RNA (siRNA) targeting both of the MEST isoforms was applied to silence MEST expression in extravillous explants and HTR8/SVneo cells. Cell invasion and migration were assessed using the Matrigel invasion, Transwell migration assay and the xCELLigence system. Promoter DNA methylation was examined using bisulfite-sequencing polymerase chain reaction (BSP).Results: MEST protein was highly expressed in EVTs in the first trimester placenta and in the invasive EVT cell lines HTR-8/Svneo and HPT-8. Weak MEST expression was found in cytotrophoblasts (CTBs) and the choriocarcinoma-derived CTB cell line JEG-3. The specific siRNA knockdown of MEST expression significantly reduced HTR-8/Svneo cell invasion and migration as well as extravillous explant outgrowth, which were associated with the downregulation of Twist, N-cadherin and Vimentin. Decreased MEST protein expression with isoform 2 promoter hypermethylation was observed in the placentas of missed abortions, suggesting a possible pathological mechanism of missed abortion.Conclusions: Suppressed expression of MEST was associated with its isoform 2 promoter hypermethylation ex vivo placenta tissues and in vitro cultured EVT cell lines. The present results provide a possible pathological mechanism of missed abortion. (C) 2016 Elsevier Ltd. All rights reserved.