多柔比星白蛋白纳米粒的制备及其对肝癌细胞Bel-7404的杀伤作用

Preparation of DOX / BSANP and its Killing Effect on Liver Cancer Cell Bel-7404

制备多柔比星白蛋白纳米粒(DOX/BSANP),观察其药物缓释规律,探讨其对肝癌细胞的杀伤作用。去溶剂-化学交联法制备DOX/BSANP载药纳米粒;透射电镜观察其形态及粒径;分光光度法测定包封率及载药率;体外药物释放实验考察缓释特性;MTT法及FCM观察其对肝癌细胞的体外杀伤作用,动物实验观察其体内抑瘤效应。透射电镜下DOX/BSANP呈圆球形,分布均匀,直径约为120 nm,动态光衍射测定其水合粒径约为170 nm,包封率为82%,载药量为5%,体外药物释放实验显示,载药纳米粒中约50%的药物持续缓慢释放,可持续长达7 d;MTT实验显示DOX/BSANP能显著抑制细胞的增殖,FCM检测显示其能诱导肿瘤细胞凋亡;体内抑瘤实验显示,载药纳米粒的疗效优于DOX原药的疗效。成功制备多柔比星白蛋白纳米粒。体内外实验证明DOX/BSANP纳米粒可有效抑制肝癌细胞Bel-7404的生长。

To prepare DOX/BSANP, and to observe its antitumor effect on liver cancer cells in vitro and in vivo, DOX/BSANP drug loading nano-particle was prepared by desolvation-chemical crosslinking method. The characters of DOX/BSANPs such as morphology, drug loading efficiency, drug entrapment efficiency and drug release rate in vitro were investigated. The chemotherapy effects of the DOX/BSANPs on liver cells in vitro were evaluated by M3~I＂ assay and flow cytometry. The liver tumor xenograft model was estab- lished on nude mice to evaluate the antitumor effect in vivo. Tumor volumes and histological analysis were performed after treating by different methods. Under transmission electron microscope, DOX/BSANP looked spherical, and uniformly distributed, diameter was about 120 nm, hydrated particle size was about 170 nm determined by dynamic light diffraction, envelopment rate was 82% and drug loading rate was 5%. In vitro drug release experiment showed that about 50% drug in drug loading nano-particle could be released continuously and slowly for 7 days. MTY test showed that DOX/BSANP could significantly inhibit cell proliferation. FCM test showed that it could induce apoptosis of tumor cells. In vivo experiments showed that therapeutic effect of drug loading nano-particle was superior to that of nude DOX. DOX/BSANP was successfully prepared. Both in vitro and in vivo experiments showed that DOX/ BSANP could effectively inhibit the growth of liver cancer cell Bel-7404.