代谢应激性自噬来源exosomes携microRNAs簇调控DLL4/Notch-VEGF通路对肝癌血管新生的作用及机制研究

肝癌是广西高发的恶性肿瘤，其多血管、高代谢使抗血管新生成为肝癌治疗的重要策略，也使代谢应激性自噬现象贯穿肝癌发展始终。DLL4/Notch-VEGF通路是应激状态影响血管新生的关键通路，明确microRNAs调控通路的作用机制对抗血管治疗具有重要意义。前期研究发现肝癌细胞代谢应激性自噬分泌液能促内皮细胞成管，基因高通量分析和细胞生物验证分泌液中外泌体的microRNAs表达，发现上调表达一组microRNAs簇与DLL4/Notch-VEGF通路密切相关。据此推测外泌体携microRNAs簇协同调控DLL4/Notch-VEGF通路在肝癌代谢应激血管新生中发挥重要作用，但机制未明。因此，本项目拟构建肝癌代谢应激性自噬模型，采用荧光标记示踪，运用体内/外实验和多种临床样本,阐明外泌体和microRNAs簇对血管新生影响及DLL4/Notch-VEGF通路调控机制，为肝癌抗血管治疗提供新思路。

Hepatocellular carcinoma is a highly-proliferated malignant tumor in Guangxi Zhuang Autonomous Region. Its multiple blood vessels and high metabolism make anti-angiogenesis an important strategy for the treatment of Hepatocellular carcinoma, as well as the phenomenon of metabolic stress autophagy throughout the development of Hepatocellular carcinoma. The DLL4 / Notch-VEGF pathway is a key pathway that affects angiogenesis under stress, and it is of great significance to clarify the regulatory mechanism of microRNAs on the pathway against vascular therapy. Previous studies have found that metabolic stress-induced autophagy secretions from hepatocellular carcinoma cells can induce endothelial cells to form tubes, high-throughput gene analysis and cell biological verification of exosomal microRNAs expression in secretions, and found that a group of microRNAs clustering is closely related to the DLL4/Notch-VEGF pathway. Based on this, it is speculated that exosomes and the microRNAs clustering coordinately regulate the DLL4/ Notch-VEGF pathway to play an important role in angiogenesis of Metabolic induced stress in Hepatocellular carcinoma, but the mechanism is unknown. Therefore, this project intends to construct a model of autophagy induced by metabolic stress in hepatocellular carcinoma, using fluorescent labeling tracing, using in vitro and in vivo experiments and a variety of clinical samples to clarify the effects of exosomes and the microRNAs clustering on angiogenesis and the DLL4 / Notch-VEGF pathway.The regulatory mechanism provides new ideas for anti-vascular treatment of Hepatocellular carcinoma.