负载EPZ6438牛血清白蛋白-壳聚糖纳米粒抑制骨肉瘤的效应  

Inhibitory effect of bovine serum albumin-chitosan nanoparticles loaded with EPZ6438 on osteosarcoma

作　　者：刘畅[1,2] 张文[1,2] 朱灿[1,2] 孙杰[1,2] 丁亦程 施勤 Liu Chang;Zhang Wen;Zhu Can;Sun Jie;Ding Yicheng;Shi Qin(First Affiliated Hospital of Soochow University,Suzhou 215006,Jiangsu Province,China;Orthopedic Institute of Soochow University,Suzhou 215123,Jiangsu Province,China;Xuzhou Medical University,Xuzhou 221000,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院,江苏省苏州市215006 [2]苏州大学骨科研究所,江苏省苏州市215123 [3]徐州医科大学,江苏省徐州市221000

出　　处：《中国组织工程研究》2024年第10期1512-1518,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(82172485),项目名称:成骨细胞Hedgehog信号持续活化调控髓源性抑制细胞参与骨质疏松症形成的作用机制及干预策略,项目负责人:施勤。

摘　　要：背景:骨肉瘤肿瘤干细胞激活最显著的转录因子是EZH2,据报道沉默EZH2可以抑制骨肉瘤细胞生长。研究证实,牛血清白蛋白-壳聚糖纳米粒是一种具有优异生物相容性和生物降解性的药物传递载体,且白蛋白载体可提供靶向肿瘤的药物递送功能。目的:探讨负载EPZ6438(EZH2抑制剂)血清白蛋白-壳聚糖纳米粒抑制骨肉瘤的效应及机制。方法:①制备未负载与负载EPZ6438的血清白蛋白-壳聚糖纳米粒,检测载EPZ6438血清白蛋白-壳聚糖纳米粒的药物包载率及药物释放率。②将MG-63细胞分4组培养,分别加入PBS(对照组)、血清白蛋白-壳聚糖纳米粒浸提液(空白纳米粒组)、EPZ6438溶液(游离药物组)及负载EPZ6438的血清白蛋白-壳聚糖纳米粒浸提液(载药纳米粒组),培养3 d后,利用流式细胞仪检测细胞凋亡、RT-PCR法检测细胞caspase3 mRNA的表达。③取12只裸鼠,通过腋下注射MG-63细胞悬液建立皮下荷瘤小鼠模型,造模成功后随机分4组干预,分别向肿瘤组织内注射生理盐水(对照组)、血清白蛋白-壳聚糖纳米粒溶液(空白纳米粒组)、EPZ6438溶液(游离药物组)及负载EPZ6438的血清白蛋白-壳聚糖纳米粒溶液(载药纳米粒组),每组3只。注射7 d后,观察肿瘤体积及肿瘤组织冰冻切片TUNEL染色。结果与结论:①载药纳米粒的药物包载率约为8.8%,该纳米粒在纯水中具有良好的药物释放效果,24 h药物释放量为(34.72±1.93)μg,72 h为(48.58±1.10)μg,120 h为(49.18±1.24)μg,168 h(50.25±1.13)μg,药物释放在120 h到达平台期,释放率约为97.9%;②与MG-63细胞培养3 d后,对照组、空白纳米粒组细胞凋亡率低于游离药物组、载药纳米粒组(P<0.001),caspase3 mRNA的表达低于游离药物组、载药纳米粒组(P<0.0001);③注射7 d后,载药纳米粒组裸鼠肿瘤体积小其他3组(P<0.05),肿瘤组织TUNEL染色阳性细胞比率高于其他3组(P<0.0001);④结果显示,负载EPZ6438的血清白蛋白-壳BACKGROUND:The most prominent transcription factor activated by tumor stem cells in osteosarcoma is EZH2,and silencing of EZH2 has been reported to inhibit osteosarcoma cell growth.Studies have confirmed that bovine serum albumin-chitosan nanoparticles are a drug delivery vector with excellent biocompatibility and biodegradability,and the albumin carrier can provide tumor-targeted drug delivery function.OBJECTIVE:To investigate the effect and mechanism of bovine serum albumin-chitosan nanoparticles loaded with EPZ6438(EZH2 inhibitor)for the treatment of osteosarcoma.METHODS:(1)Bovine serum albumin-chitosan nanoparticles loaded with and without EPZ6438 were prepared.The drug encapsulation rate and drug release rate of serum albumin-chitosan nanoparticles loaded with EPZ6438 were detected.(2)MG-63 cells were divided into four groups and added with PBS(control group),serum albumin-chitosan nanoparticle extract solution(blank nanoparticle group),EPZ6438 solution(free drug group),and serum albumin-chitosan nanoparticle extract loaded with EPZ6438(drug-loaded nanoparticle group),respectively.After 3 days of culture,cell apoptosis was detected by flow cytometry and the expression of caspase-3 mRNA was detected by RT-PCR.(3)Twelve nude mice were selected and the subcutaneous tumor-bearing mouse model was established by injecting MG-63 cell suspension under the armpit.After successful modeling,the mice were randomly divided into four groups for intervention.Normal saline(control group),serum albumin-chitosan nanoparticle solution(blank nanoparticle group),EPZ6438 solution(free drug group)and serum albumin-chitosan nanoparticle solution loaded with EPZ6438(drug-loaded nanoparticle group)were injected into tumor tissues,with three animals in each group.After 7 days of injection,the tumor volume and frozen sections of tumor tissue were observed by TUNEL staining.RESULTS AND CONCLUSION:(1)The drug encapsulation rate of the nanoparticles was about 8.8%,and the nanoparticles had a good drug release effect in pure water.The drug

关 键 词：牛血清白蛋白 壳聚糖 纳米粒 载药 骨肉瘤 细胞凋亡 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R331