TRIM24调控胶质瘤干细胞“干性”的作用机制研究

In the central nervous system, the most common primary malignant tumors are malignant gliomas. Our previous study shows that TRIM24 is closely associated with glioma malignancy. Moreover, TRIM24 promotes glioma growth and enhances chemoresistance via activating PI3K/Akt signaling. However, the mechanisms of TRIM24 in glioma development and progression are largely unveiled, especially for its roles in the stemness of glioma stem cells. Therefore, this project would be carried out as follows. At first, the glioma stem cells are isolated from surgical samples and cultured in vitro. Their tumor stemness can be identified by employing morphology, self-renewal and xenograft experiments, whilst the TRIM24 expression profiles before and after differentiation are detected. Then we take advantage of lentivirus to upregulate or downregulate TRIM24 expression in glioma stem cells, and conduct a series of in vitro and in vivo analysis to find out its biological roles in tumor stemness. Furthermore, the molecular mechanisms of TRIM24 regulation in PI3K/Akt、Wnt and Notch pathways are explored and verified by employing mutation experiment, rescue experiment, ChIP assay, dual-luciferase reporter assay and etc. This study is aimed to provide new therapeutic strategies for gliomas.

在中枢神经系统中，最常见的原发恶性肿瘤是恶性胶质细胞瘤。我们前期研究发现，TRIM24与胶质细胞瘤的恶性程度密切相关，并通过激活PI3K/Akt信号转导促进肿瘤的生长和耐药。但是，TRIM24在胶质细胞瘤发生和恶性进展中的作用机理尚不清楚，尤其是与胶质瘤干细胞“干性”的关系更是未见报道。本项目拟通过原代分离培养法获取胶质瘤干细胞，采用细胞形态观察、增殖分化、裸鼠成瘤等技术对细胞加以鉴定，并检测TRIM24在干细胞分化前后的变化情况；再利用慢病毒工具上调或下调TRIM24的表达水平，开展一系列体外和体内实验来研究TRIM24与胶质瘤干细胞“干性”的相关性；进一步探索TRIM24对胶质瘤干细胞相关的PI3K/Akt、Wnt和Notch信号通路的调控功能，并运用基因突变、表达恢复、ChIP、双荧光素酶报告基因检测等方法来验证TRIM24发挥表达调控的分子结构基础，为胶质细胞瘤的治疗提供新策略。