Promoter Hypomethylation of Maspin Inhibits Migration and Invasion of Extravillous Trophoblast Cells during Placentation.
Promoter Hypomethylation of Maspin Inhibits Migration and Invasion of Extravillous Trophoblast Cells during Placentation.
复制标题
Maspin启动子低甲基化抑制胎盘过程中绒毛外滋养层细胞的迁移和侵袭
DOI：
10.1371/journal.pone.0135359
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Wu Y
中科院分区：
文献类型：
--
作者：
Shi X;Liu H;Cao J;Liu Q;Tang G;Liu W;Liu H;Deng D;Qiao F;Wu Y
目的妊娠早期，子宫内膜外滋养细胞（EVT）侵入母体螺旋小动脉和子宫内膜。乳腺丝氨酸蛋白酶抑制剂（Maspin, SERPINB5）在调节细胞滋养层细胞的侵袭活性中起着被认为的作用。maspin基因在各种癌症中通过涉及异常胞嘧啶甲基化的表观遗传机制沉默。我们研究了maspin启动子甲基化状态对EVT细胞maspin表达和侵袭性的影响。方法采用Western blotting检测缺氧条件下EVT细胞中maspin蛋白的表达。采用细胞计数试剂盒（Cell Counting Kit-8）、流式细胞术（Flow Cytometry）和Transwell方法检测细胞的增殖能力、凋亡率和迁移侵袭能力。随后，我们用重组maspin蛋白处理细胞。采用亚硫酸盐测序PCR和甲基化特异性PCR检测缺氧/地西他滨作用下maspin启动子区的甲基化程度。最后，我们探讨了地西他滨对巨噬蛋白表达和EVT细胞侵袭性的影响。结果缺氧可有效提高EVT细胞maspin蛋白的表达，显著抑制其侵袭性。重组maspin蛋白的加入抑制了这种侵袭性。地西他滨降低了maspin启动子区域的甲基化，有效提高了maspin蛋白的表达，显著削弱了EVT细胞的迁移和侵袭性。maspin启动子的甲基化状态是影响妊娠早期EVT细胞迁移和侵袭的重要因素。甲基化状态的降低可抑制EVT细胞的迁移和侵袭，影响胎盘的发生，并可导致胎盘缺血和缺氧。
Objective Extravillous trophoblast (EVT) cells invade the endometrium and the maternal spiral arterioles during the first trimester. Mammary Serine Protease Inhibitor (Maspin, SERPINB5) plays a putative role in regulating the invasive activity of cytotrophoblasts. The maspin gene is silenced in various cancers by an epigenetic mechanism that involves aberrant cytosine methylation. We investigated the effect of the methylation status of the maspin promoter on the maspin expression and the aggressiveness of EVT cells. Methods Western blotting was used to detect the maspin protein expression in EVT cells upon hypoxia. The proliferative ability, the apoptosis rate and the migration and invasiveness were measured with Cell Counting Kit-8 assay, Flow Cytometry technology and Transwell methods. Subsequently, we treated cells with recombinant maspin protein. The methylation degree of maspin promoter region upon hypoxia/ decitabine was detected by bisulfite sequencing PCR and methylation-specific PCR. Finally, we explored the effects of decitabine on maspin protein expression and the aggressiveness of EVT cells. Results Hypoxia effectively increased maspin protein expression in EVT cells and significantly inhibited their aggressiveness. The addition of recombinant maspin protein inhibited this aggressiveness. Decitabine reduced the methylation in the maspin promoter region and effectively increased the maspin protein expression, which significantly weakened the migration and invasiveness of EVT cells. Discussion The methylation status of the maspin promoter is an important factor that affects the migration and invasion of EVT cells during early pregnancy. A decrease in the methylation status can inhibit the migration and invasion of EVT cells to affect placentation and can result in the ischemia and hypoxia of placenta.