见血封喉苷 H 诱导肝癌细胞凋亡与自噬的机制研究

项目名称： 见血封喉苷 H 诱导肝癌细胞凋亡与自噬的机制研究

项目编号： No.81460557

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 黄风迎

作者单位： 海南医学院

项目金额： 50万元

中文摘要： 见血封喉苷H（Toxicarioside H，ToxH）是我们从海南热带植物见血封喉乳汁中分离并进行了结构鉴定的新化学物。我们前期研究发现，ToxH对肝癌等多种肿瘤细胞具有较强的细胞毒活性，能够同时诱导肝癌细胞凋亡和自噬，但详细凋亡和自噬的机理、凋亡和自噬之间关系、凋亡和自噬二者是否存在共同的分子诱导机制、ToxH结合靶蛋白分子等问题仍需要深入研究。因此，本项目拟通过体外细胞培养和小鼠体内模型二个层面开展研究，利用分子生物学技术和方法了解ToxH诱导肝癌细胞凋亡和自噬的分子信号通路，确定凋亡和自噬二者之间的关系，应用磁性量子点标记技术对ToxH进行细胞定位并筛选ToxH结合的靶蛋白分子，最终利用靶分子功能阻断剂和RNA干扰等技术确认靶分子的功能，确定ToxH诱导肿瘤细胞凋亡和自噬的分子机制，为把ToxH更进一步开发成临床高效、低毒和具有自主知识产权的抗肝癌创新药物提供实验和理论依据。

中文关键词： 肝细胞癌；见血封喉苷；H；细胞凋亡；细胞自噬；分子机制

英文摘要： Toxicarioside H (ToxH) is a new cardenolide chemical compound isolated from the latex of Antiaris toxicaria in tropical Hainan Island. Its structure was elucidated by our research group and showed significant cytotoxicity against K562, SGC-7901, SMMC-7721 and HeLa cell lines in vitro by MTT method. In addition, we also found that ToxH can induce strong apoptosis and autophagy in hepatoma carcinoma cells. However, there are not any mechanism research done till now except for our group. The detail molecular mechanism of apoptosis and autophagy, the relationship between apoptosis and autophagy, the shared molecular signnaling pathway and the target protein moleculs between apoptosis and autophagy in the tumor cells induced by ToxH remains unknowed, it is thus need further more studies. In our current study, we will in vitro culture hepatoma carcinoma cells and establish in vivo tumor models to in invetigate whether apoptosis and autophagy are surely existed in hepatoma carcinoma cells induced by ToxH. In addition, we will use molecular methods to vertify the apoptotic and autophagic signaling pathways and the relationship between apoptosis and autophagy induced by ToxH. Otherwise, we will also use quantum dot (QD) method to locate the ToxH's binding-site and screen its target protein molecules in the tumor cells. Lastly, we will also use molecular blockers, RNA interference, or even gene knockout to verify the target protein molecules to ultimately prove the detail apoptitic and autophagic mechanism induced by ToxH. Our study will offer experimental data for ToxH to be developed as a novel anti-tumor compound.

英文关键词： Hepatocellular carcinoma;Toxicarioside H;Cell apoptosis;Cell autophagy;Molecular mechanism