白介素-2靶向5-氟尿苷棕榈酸酯脂质体的制备及细胞毒性研究

Preparation of 5′-Palmitoyl-5-Fluorouridine Liposomes Modified with Interleukin-2 and Its Cell Toxicity

关键词:  白介素-2  5-氟尿苷棕榈酸酯  脂质体  主动靶向  细胞毒性实验

Key words:  interleukin-2  5′-palmitoyl-5-fluorouridine  liposomes  active targeting  cell toxicity

摘要:

目的 制备了白介素-2(IL-2)靶向5-氟尿苷棕榈酸酯(5-FURP)脂质体，并进行了体外细胞毒性，初步考察其对IL-2受体高表达的肿瘤细胞的靶向作用。方法 采用逆相蒸发法制备5-FURP脂质体，通过交联剂将IL-2连接到脂质体的表面，得IL-2靶向5-FURP脂质体(IL-2-5FURP-L)；UV法测定包封率；用考马斯亮蓝结合法测定IL-2与脂质体的结合率；用MTT法测定脂质体对IL-2受体高表达的皮肤T细胞淋巴瘤Hut-102细胞的生长抑制作用。结果 IL-2-5-FURP-L的粒径为180 nm；药物平均包封率为91.3%；IL-2的结合率为56.0%；在pH7.4的释放介质中，脂质体具有缓释作用；MTT实验结果显示，IL-2-5-FURP-L呈剂量依赖性抑制肿瘤细胞的生长，72 h细胞毒性试验表明IL-2-5-FURP-L对Hut-102的杀伤作用(IC₅₀=1.04 μg·mL^-1)明显优于5-FURP-L(IC₅₀=6.11 μg·mL^-1)及5-FURP(IC₅₀=7.35 μg·mL^-1)。结论 IL-2修饰的载药脂质体具有明显的抑瘤作用和主动靶向作用。

Abstract:

OBJECTIVE To investigate the specificity of interleukin-2 (IL-2) targeting liposomes (L) containing 5 ′ -palmitoyl- 5-fluorouridine (5-FURP). METHODS Liposomes containing 5-FURP were prepared by reverse phase evaporation method. IL-2 targeting 5-FURP-L were prepared through the effect of cross linker. The entrapment efficiency was measured by UV method. The amount of IL-2 conjugated to liposomes was determined with coomassie brilliant blue binding assay. Cytotoxicity on Hut-102 cells expressing IL-2 receptors in vitro was evaluated with MTT methods. RESULTS The mean size of the liposomes was 180 nm and the entrapment efficiency was 91.3%. The coupling efficiency of IL-2 was 56.0%. In pH 7.4 PBS , 5-FURP was released in a sustained manner from the liposomes. The MTT assay suggested that the IL-2 targeting 5-FURP-L had a distinct killing effect on Hut-102 tumor cells. CONCLUSION The IL-2 modified 5-FURP-L has an active targeting function and a significant anticancer effect.