紫杉醇磁性纳米脂质体的体外细胞抑瘤效果

Depression effects of paclitaxel magnetic nanoparticle liposome on the growth of cancer cells in vitro

背景:因紫杉醇的水不溶解性质,目前多采用包埋方式载药,如被脂质体包埋或者与白蛋白结合等。目的:采用磁性纳米脂质体包埋紫杉醇,观察紫杉醇磁性纳米脂质体对几种常见肿瘤细胞株的抑制效果。设计、时间及地点:材料-细胞学体外实验,于2008-10/2009-03在解放军广州军区广州总医院药学部和医学实验科完成。材料:肺癌细胞株(SPCA-1)、卵巢癌细胞株(8910)、肝癌细胞株(HEPG-2)、黑色素瘤细胞株(B16F10)均由解放军广州军区广州总医院医学实验科提供。方法:采用共沉淀法制备Fe3O4纳米粒,同时施加超声处理减少粒子的软团聚合,然后将硬脂酸、乙醇、Tween80和紫杉醇在(73±2)℃水浴中混合搅拌至透明,迅速加入分散好的磁液,搅拌均匀形成含磁悬液,冷冻干燥后即得紫杉醇磁性纳米脂质体。复苏4种肿瘤细胞,培养至对数生长期后按1×1010L-1接种于96孔板,180μL/孔,再加入紫杉醇磁性纳米脂质体20μL,在37℃、体积分数为5%的CO2培养箱中培养48～72h。通过预实验,确定以紫杉醇磁性纳米脂质体乳液中含紫杉醇0.19g/L和0.98g/L这两个浓度进行MTT细胞毒性实验。主要观察指标:4种肿瘤细胞的MTT毒性结果。结果:两种浓度的紫杉醇磁性纳米脂质体对肺癌细胞(SPCA-1)、人卵巢癌细胞(8910)、肝癌细胞(HEPG-2)、黑色素瘤细胞(B16F10)的抑制率分别达到93.23%和93.13%、47.70%和79.12%、94.94%和99.64%、84.46%和95.03%,具有高敏感性。结论:紫杉醇磁性纳米脂质体对上述4种人肿瘤细胞具有很好的抑制作用,基本达到临床药物需要。

BACKGROUND： Paclitaxel has insolubility in water. Embedding was commonly used to carry drugs, such as liposome-embedded or albumin-combined. OBJECTIVE： To investigate the depressive effects of paclitaxel magnetic nanoparticle liposome on several common tumor ceils. DESIGN, TIME AND SETTING： The in vitro material-cytology study was performed at the Division of Pharmacy and Department of Medical Experiment of Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA from October 2008 to March 2009. MATERIALS： Lung cancer cell line （SPCA-1）, ovarian cancer cell line （8910）, hepatema carcinoma cell line （HEPG-2） and melanoma cell line （B16F10） were supplied by the Department of Medical Experiment of Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA. METHODS： Fe3O4 nanoparticles were prepared by precipitation method. Ultrasonication was used to reduce particle polymerization. Stearine, ethanol, Tween 80 and paclitaxel were mixed in water at （73±2）℃ till transparency. Subsequently, magnetic fluid was added and stirred. Following freeze drying, paclitaxel magnetic nanoparticle liposome was obtained. Four kinds of tumor cells were recovered. At logarithmic phase, cells were incubated in a 96-well plate at 1 ×10^10/L, 180 p L/well, and treated with paclitaxel magnetic nanoparticle liposome （20 pL）, and incubated in a 5% CO2 incubator at 37℃ for 48 72 hours. Following preliminary experiment, it was confirmed that paclitaxel magnetic nanoparticle liposome contained paclitaxeI at 0.19 g/L and 0.98 g/L for MTT assay. MAIN OUTCOME MEASURES： MTT toxity results of four kinds of cells. RESULTS： The inhibition ratio of the paclitaxel magnetic nanoparticle liposome of two different concentrations to the cells SPCA-1, 8910, HEPG-2, B16F10 were 93.23% and 93.13%, 47.70% and 79.12%, 94.94% and 99.64%, 84.46% and 95.03%, with hypersensitivity. CONCLUSION： Paclitaxel magnetic nanoparticle liposome really has good depressive effects on these kinds of human tumor cells, and reaches the demand of clinical medicine primitively.