系列micro-RNA荧光脂质体的制备、结构、性质和功能

A series of fluorescent micro-RMA liposomes with biological target will be prepared by combination method of supercritical CO2 and thin film evaporation. Using physical chemistry method, computer simulation and the modern detection technology, controllable self-assembly process of fluorescent liposomes will be studied. The preparation, structure and macroscopic properties and target function of micro-RNA liposomes will be revealed. The process of trans-membrane, fusion, and stimulus response switch control release of the micro-RNA will be probed in the biological system of breast tumor cells and normal cells. The relationship between the microstructure, properties and function will be explored. The interaction between the micro-RMA liposomes and biological molecules will be revealed. The interaction between the micro-RNA liposomes and living cells, microstructure, and the relationship between the macroscopic properties and microscopic structure and function will be discussed. The targeting and tumor suppressors influence on the microstructure will be uncovered. The research results will provide important information for tumor treatment, new targets for the early diagnosis of malignant tumor.

本项目采用超临界CO2-薄膜蒸发法制备具有不同微观结构的生物靶向的micro-RNA荧光脂质体。采用物理化学的研究方法、热力学的研究手段、计算机模拟和现代测试技术，探讨这些荧光脂质体可控自组装过程及其调控规律；研究不同结构不同功能荧光micro-RNA脂质体的制备、结构、宏观性质和靶向功能。以乳腺癌的肿瘤细胞和正常细胞为研究对象，荧光脂质体将micro-RNA运送到治疗干预靶点，通过micro-RNA脂质体与目标膜发生的跨膜、刺激响应开关控制释放micro-RNA等过程，研究micro-RNA荧光脂质体与活体细胞之间相互作用、宏观性质与微观结构和功能之间的关系，揭示micro-RMA脂质体的微观结构对其靶向性和生物利用度的影响及其规律。该研究的结果将为肿瘤治疗提供重要信息，为恶性肿瘤的早期诊断提供新的靶标。

恶性肿瘤的发生与发展是癌基因、抑癌基因共同作用的结果，这是一个多因素、多步骤的复杂过程。目前用于治疗肿瘤的合成药物的致命缺点是缺乏对肿瘤细胞的靶向性，在用药后不但不能使药物富集于肿瘤细胞内充分发挥其生物利用度，而且这些药物在抑制肿瘤细胞的同时杀死正常细胞，导致严重副作用。因此，寻找具有主动靶向多功能的药物载体仍然是人类亟待解决的关键问题。.本项目以探讨具有主动靶向和示踪特性的micro-RNA脂质体为目标，建立一系列新的相关研究方法，主要包括：制备micro-RNA脂质体的薄膜-超临界CO2（M-scCO2）方法；均匀分散聚合PDA于micro-RNA脂质体表面的方法；利用荧光衰减曲线获取micro-RNA脂质体表面结构信息的方法。首次采用M-scCO2方法成功将叶酸配体和PDA与脂质体结合，将Micro-RNA、PDA和多西紫杉醇负载于脂质体，制备了叶酸/PDA脂质体，Micro-RNA脂质体，多西紫杉醇/siRNA/PDA脂质体等。这些脂质体药物载体具备主动靶向给药和荧光示踪的特性。探讨了这些脂质体的表面结构，宏观特性、靶向功能和细胞毒性。