BAP31调节膀胱癌干细胞干性及其相关机制研究

项目名称： BAP31调节膀胱癌干细胞干性及其相关机制研究

项目编号： No.81502582

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 黄永业

作者单位： 东北大学

项目金额： 18万元

中文摘要： 越来越多研究表明，癌症干细胞是导致癌症难以治愈的根本原因。BAP31是一个与细胞凋亡、蛋白裂解相关的细胞因子，在干细胞分化、癌症发生中发挥重要作用。本项目拟研究BAP31在膀胱癌干细胞中的功能作用，探索其影响癌症干细胞分化的分子机制。我们采用shRNA技术，在膀胱癌细胞UC5中对BAP31表达进行稳定敲低，并运用流式细胞术分选膀胱癌干细胞。在BAP31敲低癌症干细胞中，进行自我更新、药物抗性以及侵袭转移能力等的测定。将BAP31敲低癌症干细胞植入NOD/SCID小鼠后检测体内成瘤情况，研究BAP31基因对癌症干细胞体内致瘤能力的影响。同时，分别运用Crispr-Cas9基因编辑技术和基因过表达技术在UC5中进行BAP31敲除和过表达，验证BAP31在癌症干细胞中的作用。本项目将阐明BAP31对膀胱癌干细胞分化的影响，为癌症发病机制研究及有效治疗方法的提出提供试验依据。

中文关键词： 膀胱癌；B细胞受体相关蛋白31；干性；上皮间充质转换；癌症干细胞

英文摘要： Bladder cancer is one of the most common malignancy in the world. However, the progress in bladder cancer management has been modest over the past decades. More and more reports show that there is a small sum of cells in the tumor tissue named cancer stem cells and these cells are probably accounted for the metastasis and relapse of cancer. B cell receptor associated protein 31 (BAP31) is an important cytokine, which is associated with apoptosis and protein pyrolysis. BAP31 also play a key role in regulating hESC stemness and survival through control of EpCAM expression. Furthermore, many reports have showed that BAP31 exerts its function in many types of cancers. In this study, we try to focus on the biological function of BAP31 in bladder cancer stem cells, exploiting its potential role in regulating the differentiation of tumor stem cells. The expression of BAP31 in bladder cancer cell line UC5 is stable knockdown by shRNA, and the bladder cancer stem cells are then sorted through flow cytometry. Tumor stem cell related gene expression, drug resistance and the ability of invasive and metastasis are investigated in BAP31 knockdown tumor stem cells. To determine the influence of BAP31 on the ability of in vivo tumorigenesis of tumor stem cells, NOD/SCID mice are used. After BAP31 knockdown bladder cancer stem cells being implanted into these mice, the tumor size is assessed every week. Meanwhile, in order to confirm the results harvested from BAP31 knockdown cells, studies are also performed in BAP31 knockout cells generated by Crispr-Cas9 technique and BAP31 over-expression cells, respectively. This project tries to unlock the role of BAP31 in the differentiation of tumor stem cells and enriches the molecular biology knowledge for cancer treatment.

英文关键词： bladder cancer;B-cell receptor associated protein 31;stemness;epithelial-mesenchymal transition;cancer stem cells