精准靶向CD24高表达三阴性乳腺癌细胞纳米递送系统的制备及体外验证  

Preparation and in vitro validation of nanotransmission system for accurate targeting of CD24 highexpression triple negative breast cancer cells

作　　者：周瑜清 李金穗[2,3] 陈宇 陈茂山 侯令密[2,3] 王东生[4] 蒲卢兰 邓世山[2] 周方方 Zhou Yuqing;Li Jinsui;Chen Yu;Chen Maoshan;Hou Lingmi;Wang Dongsheng;Pu Lulan;Deng Shishan;Zhou Fangfang(Department of Anatomy,School of Basic Medicine and Forensics,North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Biological Targeting Laboratory of Breast Cancer,Academician(Expert)Workstation,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Breast and Thyroid Surgery,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Breast and Thyroid Surgery,Affiliated Suining Central Hospital of Chongqing Medical University,Suining 629000,Sichuan Province,China)

机构地区：[1]川北医学院基础医学与法医学院解剖教研室,四川省南充市637000 [2]川北医学院附属医院乳腺癌生物靶向研究室院士工作站,四川省南充市637000 [3]川北医学院附属医院甲状腺乳腺外科,四川省南充市637000 [4]重庆医科大学附属遂宁市中心医院内分泌乳腺外科,四川省遂宁市629000

出　　处：《中国组织工程研究》2023年第25期4013-4019,共7页Chinese Journal of Tissue Engineering Research

基　　金：南充市2020年市校科技战略合作专项(川北医学院)资金(20SXCXTD0001),项目负责人:侯令密;四川省中医药管理局科研项目(2020JC0055),项目负责人:侯令密;南充市科技局市校科技战略合作项目(20SXQT0052),项目负责人:周方方;川北医学院2022年开放创新实验项目(SY22-060),项目负责人:周方方;川北医学院附属医院2022年科研发展计划项目(2022JC004),项目负责人:李金穗。

摘　　要：背景:纳米技术与医学的结合在选择性靶向及治疗肿瘤领域开辟了一条全新的道路,通过应用纳米载体确保了装载药物的靶向递送与稳定性,增强了细胞摄取和生物相容性。目的:制备一种谷胱甘肽和pH值双响应性CD24适配体修饰的载铁死亡激动剂Erastin和双质粒的纳米药物递送系统PC(coBP)Ca,探讨其在体外对三阴性乳腺癌中CD24高表达细胞的精准靶向性和可行性。方法:PEG-CAPDB经过多次聚合反应生成纳米微球骨架COOH-PEG-CPADB-[co BMA co PDSMA],再通过自组装合成内载铁死亡激动剂Erastin、1-2M2ge化合物、NF2敲除质粒和YAP过表达质粒形成PC(coBP),利用SELEX技术筛选CD24适配体,进一步修饰PC(coBP),最终形成载药纳米微球PC(coBP)Ca。检测PC(coBP)Ca表面CD24适配体修饰、对谷胱甘肽和pH值的敏感性、逃逸免疫细胞吞噬的性能及靶向性能、逃逸溶酶体吞噬和药物突释性能及对吞噬细胞吞噬作用的影响。结果与结论:PC(coBP)Ca的平均粒径为(141.11±13.43)nm,平均多分散性指数为0.353±0.074,平均Zeta电位显示良好分散性;PC(coBP)Ca中铁死亡激动剂Erastin的平均载药量为(23.34±2.45)%、包封率为(90.24±3.11)%;PC(coBP)Ca在谷胱甘肽和pH值双重响应作用下的4 h释放量约为90%,可有效逃避免疫细胞吞噬精准靶向CD24高表达三阴乳腺癌细胞,逃逸溶酶体,达到药物突释,激活铁死亡途径和促进吞噬细胞吞噬。实验成功合成载药纳米微球PC(coBP)Ca具备精准、有效、安全和低毒性的特点,有望成为三阴乳腺癌中CD24高表达细胞靶向纳米药物递送系统。BACKGROUND:The combination of nanotechnology and medicine has opened up a new path in the field of selective targeting and tumor treatment.Through the application of nano carriers,the targeted delivery and stability of loaded drugs are ensured,and the cell uptake and biocompatibility are enhanced.OBJECTIVE:To prepare a glutathione and pH double-responsive CD24 aptamer-modified nanodrug delivery system PC(coBP)Ca for ferroptosis agonists Erastin and biplasmids and to investigate its precise targeting and feasibility of CD24 highly expressed cells in triple negative breast cancer in vitro.METHODS:PEG-CAPDB underwent multiple polymerization reactions to form a nanosphere skeleton called:COOH-PEG-CPADB-[co BMA co PDSMA],and then PC(coBP)formation by self-assembling synthesis of the internally loaded ferroptosis agonist Erastin,1-2M2ge compound,NF2 knockout plasmid and YAP overexpressed plasmid.SELEX technology was used to screen CD24 aptamers to further modify PC(CoBP).Nano-drug delivery system PC(coBP)Ca was eventually formed.The effects of CD24 aptamer modification on the surface of PC(coBP)Ca,sensitivity to glutathione and pH,phagocytosis and targeting properties of immune cells escaping,lysosomal escaping phagocytosis and drug burst release properties,and phagocytosis of phagocytes were detected.RESULTS AND CONCLUSION:The average particle size of PC(coBP)Ca was(141.11±13.43)nm.The average polydispersity index was(0.353±0.074).The average Zeta potential showed good dispersibility.The average drug loading of Erastin was(23.34±2.45)%.The encapsulation rate of PC(coBP)Ca was(90.24±3.11)%,and the 4-hour release was about 90%under the dual response system of glutathione and pH,which could effectively evade immune cell phagocytosis to accurately target CD24 highly expressed triple negative breast cancer,escape lysosomes,then achieve drug release,which activates the ferroptosis pathway and promotes phagocyte phagocytosis.The successful synthesis of nano-delivery system PC(coBP)Ca has the characteristics of precision,

关 键 词：三阴乳腺癌 纳米递送系统 铁死亡 CD24 靶向给药 谷胱甘肽 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R979.1