天然活性分子Isatin抗神经母细胞瘤转移的作用及分子机制研究

项目名称： 天然活性分子Isatin抗神经母细胞瘤转移的作用及分子机制研究

项目编号： No.81472542

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 侯琳

作者单位： 青岛大学

项目金额： 70万元

中文摘要： 早期转移是神经母细胞瘤（NB）的重要特征，是NB患儿的主要死因，故控制转移是决定预后的关键。Isatin是存在于动植物中的天然活性分子。申请者前期研究证实Isatin有抗肿瘤活性（Eur J P 2008，2013），之后发现其能明显抑制NB转移，且能下调PI3K/Akt通路的p-Akt，据此推测Isatin可能通过PI3K/Akt通路抗NB转移。本项目拟利用NB转移瘤细胞系和可视化动物模型明确Isatin抗NB转移的作用，并探讨其分子机制：①研究其对PI3K/Akt信号通路的影响；②应用siRNA和PI3K抑制剂下调PI3K验证之；③采用亲和标记法结合蛋白飞行质谱技术探究其作用靶点，即是否通过与膜上受体结合调控该通路；④借助磷酸化蛋白芯片结合qPCR芯片，进一步筛选Isatin抑制NB转移的关键分子。本研究旨在阐明Isatin抗NB转移的作用及分子机制，以期发现NB转移治疗的潜在靶点。

中文关键词： 神经母细胞瘤；转移；吲哚-2；3-二酮；PI3K/Akt；分子机制

英文摘要： Neuroblastoma (NB) is one of the most common solid tumors in childhood that accounts for 15% of all pediatric cancer-related deaths. It is well-known that early metastasis is the major cause of the poor outcome and death of NB. However, there is no accepted therapeutics for NB metastasis due to the lack of knowledge about its molecular mechanisms. Isatin, an endogenous indole in mammalian tissues and ?uids, possesses a wide range of biological activities. Our recent studies have demonstrated that Isatin has an anti-tumor effect（Eur J P 2008，2013）. In our preliminary study, we are excited to find that Isatin could strongly inhibit the metastatic of NB, and this effect might be related to the PI3K/Akt pathway. The hypothesis of our proposal is that Isatin may have an anti-metastasis effect on NB through the PI3K/Akt pathway. The hypothesis will be tested both in cultured cells in vitro and in mice in vivo by integrative approaches. The signal molecules involved in PI3K/Akt pathway will be analyzed by fluorescence quantity RT-PCR and Western Blot. The siRNA and PI3K inhibitor will be applied to determine the effect of Isatin on PI3K/Akt pathway. The targeting molecules of Isatin will be further identified by the affinity labeling and proteins flight mass spectrometry technology. The other potential molecules that may be involved in Isatin-mediated effect on NB metastasis will be screened by phosphoralated protein chip combined qPCR chip technology. Currently, metastasis is still a well-known key challenge of NB. The proposed study, if it is successful, will identify a novel modality, as well as its-related molecular mechanisms, that could be used in the prevention and treatment of NB metastasis.

英文关键词： neuroblastoma;metastasis;isatin;PI3K/Akt;molecular mechanisms