Interaction between NF-κB and AP-1 and their intracellular localization at labor in human late pregnant myometrial cells in vivo and in vitro.
Interaction between NF-κB and AP-1 and their intracellular localization at labor in human late pregnant myometrial cells in vivo and in vitro.
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DOI：
10.1097/md.0000000000012494
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发表时间：
2018-09
期刊：
影响因子：
1.6
通讯作者：
Zhang W
中科院分区：
文献类型：
--
作者：
Peng Q;Liu Y;Dong M;Xu F;Huang J;Chen J;Li X;Zhang J;Zhang W
在发达国家，早产（PTB）是仅次于先天性异常的最重要的新生儿发病和死亡原因。据报道，NF-κB和AP-1在分娩启动中起重要作用。然而，这两种分子在分娩启动过程中的相互作用关系尚未见报道。本研究旨在探讨人妊娠晚期子宫肌瘤细胞（HLPMCs）产程中NF-κB和AP-1的相互作用及其细胞内易位。采用免疫共沉淀（Co-IP）、免疫印迹（Western blot）、免疫组化（IHC）、免疫细胞荧光（ICF）等技术探讨NF-κB与AP-1的相互作用及其在分娩前后细胞内定位的变化。自然分娩组NF-κBp65和AP-1（c-jun）蛋白表达水平明显高于非分娩组。Pearson相关分析显示NF-κBp65蛋白表达与AP-1（c-jun）呈正相关。在自然分娩组和非分娩组的hlpmc中均发现了这两个分子之间的相互作用，在原代培养的hlpmc中，在神经介质B （NMB）刺激前后也发现了这两个分子之间的相互作用。NF-κBp65和AP-1（c-jun）在分娩开始或NMB刺激前主要定位于细胞质，在分娩开始和NMB刺激时转移到细胞核中。这些结果表明，NF-κBp65和AP-1（c-jun）蛋白表达上调、相互作用增强以及它们向细胞核的易位可能与分娩起始有关。
Preterm birth (PTB) is the most important cause of neonatal morbidity and mortality next to congenital anomalies in the developed world. NF-κB and AP-1 were reported to play an important role in parturition initiation. However, the interaction relationship between the 2 molecules in labor initiation has not yet been reported. This study aimed to investigate the interaction between NF-κB and AP-1 and their intracellular translocation during labor in human late pregnant myometrial cells (HLPMCs). Co-immunoprecipitation (Co-IP), Western blot analysis, immunohistochemistry (IHC), and immunocytofluorescence (ICF) techniques were applied to explore the interaction between NF-κB and AP-1 and the alteration in their intracellular localization before and after labor onset. The protein expression levels of NF-κBp65 and AP-1(c-jun) in the natural labor group were observed significantly higher than that in the non-labor group. Pearson's correlation analysis showed a positive correlation between the protein expression of NF-κBp65 and AP-1(c-jun). Interactions were found between the 2 molecules in HLPMCs both in natural labor and non-labor group and were also found in primary culture HLPMCs before and after neuromedin B (NMB) stimulation. NF-κBp65 and AP-1(c-jun) were localized mainly in the cytoplasm before labor onset or NMB stimulation and were translocated into the nucleus upon labor initiation and NMB stimulation. These results demonstrated that upregulated protein expression of NF-κBp65 and AP-1(c-jun), the enhanced interaction between the 2 molecules, and their translocation to nucleus might be correlated to labor initiation.