TNFR1/ANXA1相互作用对胶质瘤增殖和侵袭的影响

The inflammatory microenvironment plays an important role in various stages of the development of gliomas. This project will combine protein interactions and gene therapy, tumor necrosis factor receptor 1 (TNFR1) activated by inflammatory cytokine TNF alpha interacted with membrane protein A1 (ANXA1), promote glioma cell proliferation and invasion, which was regarded as a breakthrough. This study focuses the mechanisms of inflammatory microenvironment in the occurrence and development of glioma. Firstly, in the tissue level, analyze the relationship between the expression TNFR1/ANXA1 and gliomas grades, clinicopathologic variables and patients’ survival. Secondly, in vitro, we will observe the influence on the expression of p-ANXA1，MEK1，ERK and cell proliferation and invasion after interfering the combination of TNFR1 and ANXA1, clarify the regulating mechanism of the interactions of TNFR1/ANXA1 in gliomas. Finally, in vivo, we study the biological role of the interactions of TNFR1/ANXA1 in glioma occurrence by transplanting glioma cells which over-expressed or knock down of TNFR1/ANXA1 into the brain of nude mouse, or by injecting into the gliomas nude mouse model with lentiviral which could target intervene the expression of TNFR1/ANXA1. Completing the present study will provide the reliable experimental and theroretical basis for gliomas treatment.

炎症微环境与胶质瘤的发生密切相关。本项目将蛋白质相互作用和基因治疗结合起来，从肿瘤坏死因子受体1(tumor necrosis factor receptor 1，TNFR1）被炎症因子TNFα激活后，与膜联蛋白A1（Annexin A1，ANXA1）形成复合物，促进细胞增殖、侵袭入手，研究胶质瘤的发病机制。首先在组织水平，研究TNFR1、ANXA1表达与临床病理及预后的关系；其次在细胞水平，研究炎症微环境下，干预TNFR1/ANXA1对p-ANXA1、MEK1、ERK及细胞增殖、侵袭的影响，阐明TNFR1/ANXA1在胶质瘤发生中的调控机制；最后在整体水平，将干预TNFR1/ANXA1表达的胶质瘤细胞移植入裸鼠脑内，或将靶向干预TNFR1/ANXA1的慢病毒载体注入裸鼠胶质瘤模型脑内，观察对胶质瘤形成的影响。课题的完成将为胶质瘤的治疗提供可靠的实验基础和理论依据。

肿瘤炎症微环境在胶质瘤发展的各阶段起到重要作用。项目拟验证在胶质瘤炎症微环境中肿瘤坏死因子受体1（TNFR1）与膜联蛋白A1（ANXA1）相互作用，使ANXA1有可能被由TNFR1招募的PKC磷酸化，从而通过调节下游信号通路促使胶质瘤细胞发生增殖、侵袭。项目首先在组织水平发现TNFR1、ANXA1在胶质瘤组织中的表达均随肿瘤恶性程度的增加而逐渐增高，统计分析显示二者均与增殖指数Ki-67呈正相关，且高表达TNFR1、ANXA1与患者的不良预后有关；其次在细胞水平发现TNFR1、ANXA1在胶质瘤细胞中存在相互作用并可诱导ANXA1酪氨酸磷酸化；在血清饥饿再释放诱导胶质瘤细胞增殖过程中，TNFR1、ANXA1表达均明显增高；TNF-α刺激诱导的炎症微环境中， ANXA1发生核转位，从而调节下游PI3K-Akt信号通路；最后通过转染ANXA1小干扰RNA发现，下调ANXA1表达可以抑制由TNF-α刺激介导的胶质瘤细胞增殖及侵袭。综上所述，项目的研究结果揭示在胶质瘤炎症微环境中，TNFR1能与ANXA1相互结合形成复合物，促使ANXA1入核发生酪氨酸磷酸化，从而调节下游PI3K-Akt信号通路促进胶质瘤细胞增殖、侵袭。.按照合同约定，课题研究成果以论文形式提交。在该基金共资助下，目前已发表论文3篇（其中SCI收录2篇），协助培养硕士研究生2名。