F806诱导F-Actin重构抗食管鳞癌分子机制研究

我们以往研究发现，大环内酯类分子F806具有显著的体内外抗食管鳞癌活性，可显著抑制癌细胞的生长、移动、侵袭与粘附，诱导癌细胞凋亡。近年我们进一步研究发现，F806作用于食管鳞癌细胞，细胞微丝骨架F-Actin发生明显回缩重构并伴随小泡生成，提示细胞微丝骨架可能是F806发挥其功能的关键效应部位。然而，F806究竟直接作用于癌细胞的哪一蛋白分子，是细胞微丝骨架某相关蛋白，还是微丝骨架蛋白本身，诱发F-Actin回缩重构，这一重要科学问题尚不清楚。为此，本项目拟采用F806偶联蛋白亲和层析、蛋白质谱、计算机分子对接、蛋白关键氨基酸定点突变、激光共聚焦活细胞定位、实时细胞功能分析以及动物活体成像等实验技术，鉴定F806抗食管鳞癌直接作用靶蛋白，研究F806与该靶蛋白相互作用特征，深刻揭示F806诱导F-Actin回缩重构，抗食管鳞癌分子机制，为探索F806作为抗食管鳞癌先导药物提供科学理论依据。

Our previous study found that macrolide analog F806 exhibited potent activity against human esophageal squamous cell carcinoma in vivo and in vitro. Furthermore, F806 inhibited various cell biological activity including cell growth, migration, invasion and adhesion, ultimately resulting in cell apoptosis. Intrestingly, our recent results show the rearrangement and small vesicles of F-Actin cytoskeleton in F806-treated esophageal squamous cell carcinoma cells, hinting F-Actin as the key effector of F806. However, an important scientific problem about the direct targeted molecule of F806 inducing F-Actin rearrangement remains unkwown in esophageal squamous cell carcinoma cells. It may be F-Actin related protein, or F-Actin itself. Therefore, this project plans to identify the direct targeted protein of F806, investigate the characteristics of interaction between F806 and targeted protein, and profoundly reveal the molecular mechanism of F806-induced F-Actin rearrangement against esophageal squamous cell carcinoma by multiple methods such as affinity chromatography, proteome analysis, computer molecular docking, site-directed mutagenesis, the laser confocal imaging living cells, real-time cell function analysis and In-vivo imaging system. Eventually, it will provide preliminary scientific theory basis for exploring F806 as a novel compound against esophageal squamous cell carcinoma.