The cAMP-responsive element binding protein (CREB) transcription factor regulates furin expression during human trophoblast syncytialization
The cAMP-responsive element binding protein (CREB) transcription factor regulates furin expression during human trophoblast syncytialization
复制标题
cAMP 反应元件结合蛋白 (CREB) 转录因子在人滋养层合胞化过程中调节弗林蛋白酶的表达
DOI：
10.1016/j.placenta.2014.07.017
复制
发表时间：
2014-11-01
期刊：
影响因子：
3.8
通讯作者：
Wang, H.
中科院分区：
文献类型：
--
作者：
Zhou, Z.;Wang, R.;Wang, H.
简介：多核合胞体滋养层是由细胞滋养层细胞融合形成和维持的，具有多种功能，以确保成功妊娠。方法：采用绒毛膜癌BeWo细胞诱导融合和原代培养的足月细胞滋养层细胞自发融合模型，采用实时荧光定量PCR、Western blotting和免疫荧光检测furin的表达。通过荧光素酶报告基因分析、截短突变体分析、定点突变和ChIP等方法鉴定了调控FUR基因启动子的关键转录因子和关键反应元件。我们证明，在细胞滋养层细胞自发融合和Forskolin诱导的BeWo细胞融合过程中，cAMP/PKA信号通路显著刺激FUR mRNA的水平，cAMP反应元件结合蛋白（CREB）被证明是在Forskolin诱导的BeWo细胞融合过程中调控FUR P1启动子的关键转录因子，并进一步鉴定了P1启动子中的两个关键cAMP反应元件（克雷斯）。最后，我们证明CREB介导了内源性弗林蛋白酶激活，并且CREB siRNA减弱了BeWo细胞中毛喉素诱导的弗林蛋白酶表达和细胞融合。讨论：这提供了滋养层细胞融合期间弗林蛋白酶上游调节因子的第一个证据。结论：上述结果表明，FUR转录激活CREB依赖性刺激的FUR P1启动子在人滋养层合胞。(C)2014爱思唯尔有限公司版权所有。
Introduction: The multinucleated syncytiotrophoblast is formed and maintained by cytotrophoblast cell fusion and serves multiple functions to ensure a successful pregnancy. We have previously reported that the proprotein convertase furin is required for trophoblast syncytialization by processing type 1 insulin-like growth factor receptor (IGF1R).Methods: Utilizing trophoblast cell fusion models including induced fusion of choriocarcinoma BeWo cells and spontaneous fusion of primary cultured term cytotrophoblast cells, the expression of furin was evaluated by quantitative real-time PCR, Western blotting and immunofluorescence. The key transcription factor regulating the FUR gene promoter and critical responsive elements were identified by luciferase reporter assays, truncated mutants analysis, site-directed mutagenesis and ChIP.Results: We demonstrated that the levels of FUR mRNA were significantly stimulated by cAMP/PKA signaling pathway during spontaneous fusion of cytotrophoblast cells and forskolin-induced fusion of BeWo cells, cAMP-responsive element binding protein (CREB) was proven to be the key transcription factor which regulated the FUR P1 promoter during forskolin-induced BeWo cell fusion, and two critical cAMP-responsive elements (CREs) in the P1 promoter were further identified. Finally, we showed that CREB mediated endogenous furin activation and that CREB siRNA attenuated forskolin-induced furin expression and cell fusion in BeWo cells.Discussion: This provides the first evidence of the upstream regulator of furin during trophoblast cell fusion. Conclusions: The above results suggest that the FUR transcription is activated by CREB-dependent stimulation of the FUR P1 promoter during human trophoblast syncytialization. (C) 2014 Elsevier Ltd. All rights reserved.