双酚A暴露对神经嵴干细胞良恶性分化潜能的影响

项目名称： 双酚A暴露对神经嵴干细胞良恶性分化潜能的影响

项目编号： No.81201522

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 肿瘤学1

项目作者： 朱海涛

作者单位： 复旦大学

项目金额： 23万元

中文摘要： 神经母细胞瘤(NB)为婴幼儿最常见恶性实体肿瘤之一。胚胎期神经嵴干细胞（NC）向外周交感神经元的良性分化受阻及向肿瘤干细胞恶性转化是NB发生的关键过程，诱因及机制尚不明确。环境内分泌干扰物（EED）是新近发现威胁人类健康的致病因素。申请者前期研究发现双酚A（BPA）等EED通过β-雌激素受体（ER）介导可促进NB进展恶化。本课题拟体外诱导人多能性干细胞分化为NC，采用激光共聚焦显微技术观察不同浓度BPA对NC向交感神经元体外定向分化的影响；应用流式细胞分选技术、体外细胞克隆形成实验及裸鼠移植瘤模型观察BPA在NC向NB肿瘤干细胞恶性转化中的作用；应用荧光定量PCR及免疫印迹分析BPA对ERβ表达的影响，并运用RNAi等技术分析ERβ不同表达水平对BPA作用的影响。揭示BPA在NB发生中的作用及机制，阐明EED与NB发生的相关性，为NB的早期预防及相关公共卫生政策制定提供依据。

中文关键词： 神经母细胞瘤；神经嵴干细胞；分化；双酚A；雌激素受体

英文摘要： Neuroblastoma(NB) is one of the most common solid malignant tumors in children.NB derives from neural crest cells (NC) which can not be differentiated normally and spin off cancer stem cells during the embryo development. However, little is known about the risk factors and their mechanisms. Environmental endocrine disruptors (EED) are recently identified risk factors associated with various human diseases. Our previous studies have implicated the role of EED including bisphenol A (BPA) in neuroblastoma cell proliferation, multidrug resistance, migration and invasion. The estrogen receptorβ（ERβ）-dependent pathway may contribute to the underlying mechanisms. Here, we further investigated the effects of BPA exposure on both benign and malignant differentiative potential of neural crest stem cells. Normal human induced pluripotent stem cells (hIPS) were differentiated into NC in vitro. Dose-time effects of BPA on the differentiation of NC into sympathetic neurons were detected by the confocal microscopy. BPA-induced malignant differentiative potential of NC into neuroblastoma stem cells was evaluated by flow cytometry, soft agar clonogenic and tumorigenesis assays. ERβ mRNA and protein expressions were assessed by real-time PCR and western blot analysis. The association between toxic effects of BPA and ERβ expressio

英文关键词： neuroblastoma；neural crest stem cells；differentiation；bisphenol A；estrogen receptor