MTA3 Regulates Extravillous Trophoblast Invasion Through NuRD Complex.
MTA3 Regulates Extravillous Trophoblast Invasion Through NuRD Complex.
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DOI：
10.3934/medsci.2017.1.17
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发表时间：
2017
影响因子：
0.7
通讯作者：
Wang K
中科院分区：
文献类型：
--
作者：
Chen Y;Khoo SK;Leach R;Wang K
绒毛外滋养层细胞（EVT）的侵入是重塑子宫三级动脉和胎盘发育所必需的。受损的EVT侵袭可能导致胎盘相关疾病的病理学改变。转移相关蛋白3（MTA 3）是核小体重塑和去乙酰化（NuRD）复合物的亚基之一，其以组蛋白去乙酰化酶依赖的方式抑制转录。据报道，MTA 3在先兆子痫胎盘中下调，表明其在EVT侵袭中的潜在作用。在这里，我们调查的作用，MTA 3在EVT入侵通过研究其分子机制在EVT细胞。首先，我们使用免疫组织化学证实了MTA 3在人胎盘EVT细胞中的表达。然后，我们使用慢病毒介导的MTA 3短发夹RNA（shRNA）敲低EVT衍生的HTR 8/SVneo细胞中的MTA 3表达，并发现MTA 3敲低细胞的侵袭能力更高。使用定量实时PCR，我们发现与对照细胞相比，在MTA 3敲低的细胞中，侵袭相关基因基质金属蛋白酶2（MMP 2）、基质金属蛋白酶9（MMP 9）和转录因子Snail的表达更高。免疫共沉淀-Western blot显示组蛋白去乙酰化酶1（HDAC 1）与MTA 3在HTR 8/SVneo细胞中的蛋白质-蛋白质相互作用。免疫共沉淀-质谱分析进一步鉴定了71种与MTA 3相互作用的蛋白，包括NuRD亚基、异染色质蛋白、表观遗传修饰剂和转录因子。这一结果不仅表明了NuRD复合物参与了MTA 3的功能，而且也证实了在EVT中MTA 3和NuRD复合物介导的转录抑制中存在复杂的多个共同参与者。总之，我们的数据表明，MTA 3调节EVT入侵和相关基因的表达，通过在EVT的NuRD复合物。
Extravillous trophoblast (EVT) invasion is required for remodeling uterine tertiary arteries and placenta development during pregnancy. Compromised EVT invasion may contribute to the pathology of placenta-related diseases. Metastasis -associated protein 3 (MTA3) is one of the subunits of nucleosome remodeling and deacetylation (NuRD) complex that represses transcription in a histone deacetylase-dependent manner. MTA3 is reported to be down-regulated in preeclamptic placentas, suggesting its potential role in EVT invasion. Here, we investigate the role of MTA3 in EVT invasion by studying its molecular mechanisms in EVT cells. First, we confirmed MTA3 expression in the EVT cells in human placenta using immunohistochemistry. We then used lentivirus-mediated MTA3 short hairpin RNA (shRNA) to knock down MTA3 expression in EVT-derived HTR8/SVneo cells and found higher invasion capacity in MTA3 knockdown cells. Using quantitative real-time PCR, we showed higher expression of invasion-related genes matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), and transcription factor Snail in MTA3 knockdown compared with control cells. Co-immunoprecipitation-Western blot assay showed the protein-protein interaction of histone deacetylase 1 (HDAC1), a subunit of NuRD, with MTA3 in HTR8/SVneo cells. Co-immunoprecipitation-Mass spectrometry assay further identified 71 proteins interacting with MTA3, including NuRD subunits, heterochromatin proteins, epigenetics modifiers and transcription factors. This result not only indicated the involvement of NuRD complex in MTA3’s function, but also demonstrated the complicated multiple co-players in MTA3 and NuRD complex mediated transcription repression in EVT. In summary, our data demonstrates that MTA3 regulates EVT invasion and related gene expression via NuRD complex in EVT.