LHRHa靶向紫杉醇脂质体的制备及体外抗肿瘤作用

Preparation of LHRHa Paclitaxel liposomes and its anti tumor effect in vitro

摘要:

目的：制备促黄体激素释放激素类似物（Luteinizing hormone releasing hormone analogues,LHRHa)靶向紫杉醇脂质体（Paclitaxel liposomes,PTX-Lipo），研究其在体外增强紫杉醇（Paclitaxel,PTX）对卵巢癌A2780/DDP细胞的抑制作用。方法：采用薄膜超声法制备PTX-Lipo与LHRHa靶向紫杉醇脂质体（LHRHa-Paclitaxel liposomes,LHRHa-PTX-Lipo），用透射电镜考察脂质体形态；高效液相色谱法测定2种PTX-Lipo的包封率；激光共聚焦法通过卵巢癌A2780/DDP细胞对4-氟-7-硝基-2，1，3-苯并氧杂恶二唑荧光素的摄取检测来反映细胞对NBD-Lipo与NBD-LHRHa-Lipo的摄取情况；MTT法及细胞克隆形成实验检测LHRHa-PTX-Lipo体外对卵巢癌细胞的生长抑制情况。-结果：制备LHRHa-PTX-Lipo的平均粒径123.4 nm，包封率在90%以上;A2780/DDP细胞对NBD-LHRHa-Lipo组的荧光摄取明显高于NBD-Lipo组；LHRHa-PTX-Lipo对A2780/DDP细胞的生长及克隆形成抑制明显高于PTX组及PTX-Lipo组（P＜0.05）。结论：采用薄膜超声法制备的LHRHa-PTX-Lipo可使药物在靶部位聚集，增强药物对卵巢癌细胞的抑制作用。

Abstract:

Objective:To prepare LHRHa-Paclitaxel liposomes and to observe its inhibitory effect on ovarian cancer A2780/DDP cells in vitro. Methods:Paclitaxel-liposomes(PTX-Lipo) and LHRHa-Paclitaxel liposomes (LHRHa-PTX-Lipo) were prepared by means of film ultrasound and their morphology was examined with transmission electron microscope.The encapsulation rates of PTX-Lipo and LHRHa-PTX-Lipo were quantified with high performance liquid chromatography (HPLC).The cellular intake of NBD-Lipo and LHRHa-NBD-Lipo was embodied through detecting the intake of NBD fluorescein by ovarian cancer A2780/DDP cells with laser confocal microscope.The inhibitory effect of LHRHa-PTX-Lipo on ovarian cancer cells in vitro was measured by MTT assay and colony forming assay.Results:The mean diameter of LHRHa-PTX-Lipo particles was 123.4 nm with encapsulation rate being above 90%.The fluorescein intake of A2780/DDP cells was higher in NBD-LHRHa-Lipo group compared with that in NBD-Lipo group.The inhibitory effect of LHRHa-PTX-Lipo on the growth and clone formation of A2780/DDP cell was much more intense than that of PTX and PTX-Lipo(P<0.05).Conclusion:LHRHa-PTX-Lipo prepared by means of film ultrasound could aggregate drugs at targeted site thus increase the inhibitory effect of drugs on ovarian cancer cells in vitro.