MNSFβ Promotes the Proliferation and Migration of Human Extravillous Trophoblast Cells and the Villus Expression Level of MNSFβ Is Decreased in Recurrent Miscarriage Patients
MNSFβ Promotes the Proliferation and Migration of Human Extravillous Trophoblast Cells and the Villus Expression Level of MNSFβ Is Decreased in Recurrent Miscarriage Patients
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MNSFβ 促进人绒毛外滋养层细胞的增殖和迁移，反复流产患者绒毛中 MNSFβ 表达水平降低
DOI：
10.1159/000506309
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发表时间：
2020-12-16
影响因子：
2.1
通讯作者：
Wang, Jian
中科院分区：
文献类型：
--
作者：
Wang, Nan;Yang, Qian;Wang, Jian
目的：卵泡外滋养细胞（EVT）侵入母体蜕膜对妊娠的建立和维持至关重要。EVT细胞侵袭紊乱可能导致妊娠并发症，包括复发性流产（RM）。我们之前报道过单克隆非特异性抑制因子β (MNSFβ)缺乏导致小鼠早期妊娠失败，RM患者个体MNSFβ表达水平显著降低，但MNSFβ在母胎界面作用的潜在分子机制尚不清楚。因此，在本研究中，我们确定了下调MNSFβ表达对人EVT细胞活性的影响。方法：采用免疫荧光法和免疫组织化学法分别检测MNSFβ在妊娠早期人蜕膜和胎盘绒毛组织中的表达。转染MNSFβ小干扰RNA可下调MNSFβ的表达，转染重组pDsRed-MNSFβ质粒可上调MNSFβ表达。采用细胞术、划痕实验、transwell实验和FITC/PI染色分别检测HTR8/SVneo细胞的增殖、迁移、侵袭和凋亡活性。Western blot检测RM患者和正常孕妇（NP）胎盘绒毛中P53、RhoA、Bcl-2、Bax和MMP-9的表达水平以及MNSFβ和RhoA的表达水平。结果：MNSFβ蛋白在妊娠早期人绒毛和绒毛外滋养细胞中均有表达。MNSFβ表达下调显著减弱HTR8/SVneo细胞的增殖、迁移和侵袭能力，P53、RhoA、Bcl-2、Bax和MMP-9的表达水平明显降低，而MNSFβ表达上调在HTR8/SVneo细胞中呈相反作用。此外，RM患者妊娠早期胎盘绒毛组织中MNSFβ和RhoA的表达水平明显低于NP患者。结论：这些数据提示MNSFβ可能通过P53信号通路促进人EVT细胞的增殖和迁移，胎盘绒毛中MNSFβ的缺乏可能通过降低EVT的增殖和侵袭活性导致早期妊娠丢失。
Aims: The invasion of extravillous trophoblast (EVT) cells into maternal decidua is essential for the establishment and maintenance of pregnancy. Derangement of EVT cell invasion might cause pregnancy complications including recurrent miscarriage (RM). We previously reported that deficiency of monoclonal nonspecific suppressor factor beta (MNSFβ) led to the early pregnancy failure in mice and the decidual MNSFβ expression level in RM patients was significantly decreased, but the underlying molecular mechanism of the role that MNSFβ played at the maternal-fetal interface remains unclear. Thus, in the present study, we determined effects of downregulated MNSFβ expression on human EVT cell activities. Methods: The MNSFβ expression in first-trimester human decidual and placental villus tissues was detected, respectively, by immunofluorescence or immunohistochemical analyses. The MNSFβ expression level in the immortalized first-trimester human EVT cell line HTR8/SVneo was downregulated by transfecting the small interfering RNA against MNSFβ and upregulated by transfecting the recombinant pDsRed-MNSFβ plasmids. The proliferation, migration, invasion, and apoptosis activities of HTR8/SVneo cells were, respectively, determined by cytometry assay, scratch test, transwell assay, and FITC/PI staining. The expression levels of P53, RhoA, Bcl-2, Bax, and MMP-9 in HTR8/SVneo cells, as well as the expression levels of MNSFβ and RhoA in placental villi of RM patients and physically normal pregnant women (NP), were examined by Western blot analysis. Results: MNSFβ protein signals were observed in first-trimester human villus and extravillous trophoblast cells. The downregulated MNSFβ expression significantly attenuated the proliferation, migration, and invasion abilities of HTR8/SVneo cells, accompanied with the obviously decreased expression levels of P53, RhoA, Bcl-2, Bax, and MMP-9, whereas the upregulated MNSFβ expression in HTR8/SVneo cells represented the inverse effects. Furthermore, expression levels of MNSFβ and RhoA in first-trimester human placental villus tissues of RM patients were significantly decreased compared to that of NP women. Conclusion: These data suggested that MNSFβ promotes proliferation and migration of human EVT cells, probably via the P53 signaling pathway, and the deficiency of MNSFβ in placental villi might lead to early pregnancy loss by reducing proliferation and invasion activities of EVTs.