载氟尿嘧啶的肝细胞癌靶向外泌体构建与疗效评价
Construction and Efficacy Evaluation of Fluorouracil-Loading Hepatocellular Carcinoma-Targeted Exosomes
DOI: 10.12677/hjbm.2025.154079, PDF,   
作者: 饶立威, 杨 勇*：中国药科大学基础医学与临床药学学院，江苏 南京；覃虹锟, 李 睿*：重庆药友制药有限责任公司，重庆
关键词: 肝细胞癌；外泌体；药物递送；氟尿嘧啶；Hepatocellular Carcinoma； Exosomes； Drug Delivery； 5-Fluorouracil
摘要: 目的：构建一种负载氟尿嘧啶的肝细胞癌靶向外泌体，并对其疗效与安全性进行评价。方法：采用慢病毒转染法，将磷脂酰肌醇蛋白聚糖-3单克隆抗体单链可变区修饰至外泌体表面，构建肝细胞癌靶向外泌体GPC3-Exo，用体外细胞摄取实验，研究其肝细胞癌靶向能力；采用超声处理法，将氟尿嘧啶载入外泌体，得载药外泌体GPC3-Exo-5FU，用细胞毒性实验，研究其对肝细胞癌细胞的杀伤能力；最后构建肝细胞癌皮下荷瘤裸鼠模型，研究载药外泌体的体内疗效及安全性。结果：成功构建肝细胞癌靶向外泌体GPC3-Exo，用多种细胞对GPC3-Exo的靶向能力进行探究，发现GPC3-Exo可高效靶向肝细胞癌细胞；经超声载药处理，得载药外泌体GPC3-Exo-5FU，载药率为(4.96 ± 0.26)%，用肝细胞癌细胞对GPC3-Exo-5FU的细胞毒性进行评估，发现GPC3-Exo-5FU可有效抑制肝细胞癌细胞的活力；最后用肝细胞癌皮下荷瘤小鼠评价GPC3-Exo-5FU的体内药效及安全性，发现GPC3-Exo-5FU有效抑制小鼠肿瘤增殖，抗肿瘤疗效与安全性均强于游离氟尿嘧啶。结论：本研究成功构建了一种负载氟尿嘧啶的肝细胞癌靶向外泌体，在肝细胞癌治疗中表现出良好的疗效及安全性。
Abstract: Objective: This study aimed to develop a hepatocellular carcinoma (HCC)-targeted exosome system loaded with 5-fluorouracil (5-FU) and to evaluate its therapeutic efficacy and safety. Methods: Glypican-3 (GPC3)-specific single-chain variable fragment (scFv) was incorporated onto the exosome membrane via lentiviral transfection to generate GPC3-targeted exosomes (GPC3-Exo). The targeting ability of GPC3-Exo toward HCC cells was assessed using in vitro cellular uptake assays. 5-FU was loaded into GPC3-Exo by ultrasonic treatment to prepare the drug-loaded exosomes (GPC3-Exo-5FU). Cytotoxicity assays were performed to evaluate the anti-tumor activity of GPC3-Exo-5FU against HCC cells in vitro. Finally, a subcutaneous HCC xenograft nude mouse model was established to assess the in vivo therapeutic efficacy and safety of GPC3-Exo-5FU. Results: GPC3-Exo was successfully constructed and demonstrated efficient targeting toward HCC cells in various in vitro models. Following ultrasonic drug loading, GPC3-Exo-5FU was obtained with a drug-loading efficiency of 4.96% ± 0.26%. In vitro cytotoxicity assays confirmed that GPC3-Exo-5FU significantly inhibited the viability of HCC cells. In vivo studies using HCC-bearing nude mice showed that GPC3-Exo-5FU effectively suppressed tumor growth, with superior anti-tumor efficacy and safety compared to free 5-FU. Conclusion: This study successfully established a 5-FU-loaded, HCC-targeted exosome delivery system that demonstrated promising therapeutic potential and safety in the treatment of hepatocellular carcinoma.
文章引用：饶立威, 覃虹锟, 李睿, 杨勇. 载氟尿嘧啶的肝细胞癌靶向外泌体构建与疗效评价[J]. 生物医学, 2025, 15(4): 725-736. https://doi.org/10.12677/hjbm.2025.154079

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