长链非编码RNA H19 在2,3,7,8－四氯代二苯并对二恶英诱导小鼠腭裂中的作用机制

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is the representative of dioxin toxicity and it can lead to various birth defects including cleft palate. In the previous study, we found that the expression of long non-coding RNA H19 (lncRNA H19) was up-regulation in the key palatal growth of mice induced by TCDD. The TGF-β3 growth factor could decrease the mRNA expressions of lncRNA H19 in the mouse embryonic palatal mesenchymal (MEPM) cells, and we found the some genes of TGF-β/Smad pathway would changed when siRNA targeting lncRNA H19. The data suggested that there was an interaction between lncRNA H19 and TGF-β/Smad pathway. To investigate the molecular mechanism between them, we used MEPM cells culture systems in vitro, a bilateral palatal shelves organ culture model and cleft palate animal model in vivo. Using the molecular biology and histopathology technology, we will explore the interaction between lncRNA H19 and TGF-β/Smad signaling pathway, which affect on the disappearance of midline epithelial seam and the proliferation, migration, differentiation of MEPM cells during the secondary palate development of fetal mouse. The aim of this work is to reveal mechanism of lncRNA H19 in cleft palate of mouse induced by TCDD.

2,3,7,8－四氯代二苯并对二噁英（TCDD）作为二噁英毒性效应的代表性受试物可导致腭裂在内的多种出生缺陷如腭裂。我们前期工作发现TCDD诱导胎鼠腭裂组织中长链非编码RNA H19（lncRNA H19）在腭生长的关键时期表达上调；TCDD处理胎鼠腭间充质（MEPM）细胞中lncRNA H19表达上调，而TGF-b3生长因子处理后lncRNA H19的表达呈下降趋势；在MEPM细胞中siRNA干扰lncRNA H19，TGF-β/Smad信号途径中信息分子也会改变。这些资料显示lncRNA H19和TGF-β/Smad途径间存在相互作用。为阐明二者间关系，我们建立体内外腭裂模型并结合现代分子病理学技术，在胎鼠次生腭发育期，探索lncRNA H19和TGF-β3/Smad信号途径的相互作用，以及这种作用对上皮中嵴缝的消失，MEPM细胞增殖、迁移、分泌、分化等的影响，来揭示lncRNA H19在TCDD诱导的腭裂中的作用机制。

2,3,7,8－四氯代二苯并对二恶英（TCDD）作为二噁英毒性效应的代表性受试物可导致腭裂在内的多种出生缺陷。但TCDD诱导胎鼠腭裂表观遗传学机制如长链非编码RNA（long non-coding RNA H19，lncRNA H19）仍不清楚。本课题通过分子生物学、细胞生物学和现代分子病理学等方法与技术，对lncRNA H19在TCDD诱导小鼠腭裂中的作用机制进行了初步研究。申请人研究结果表明TCDD处理小鼠腭间充质（MEPM）细胞中lncRNA H19表达上调，TGF-β3处理后lncRNA H19的表达呈下降趋势；在MEPM细胞中敲除lncRNA H19，TGF-β/Smad信号途径基因也会改变，同时进一步阐明TCDD诱导胎鼠腭裂中lncRNA H19参与TGF-β/Smad信号通路中的分子机制，为研究腭裂的发生机制提供新思路。