载氟尿嘧啶聚乳酸碳纳米管复合材料对胃癌细胞株的体外杀伤效应  

Anti-tumor effect of 5-FU-PLLA-CNTs on human gastric carcinoma cell lines in vitro

作　　者：顾钧[1] 李茂岚[1] 吴向嵩[1] 吴文广[1] 张林[1] 丁琦晨[1] 杨佳华[1] 翁昊[1] 丁倩[1] 包润发[1] 束翌俊 刘颖斌[1] 

机构地区：[1]上海交通大学医学院附属新华医院普通外科普通外科实验室上海交通大学医学院胆道疾病研究所,200092

出　　处：《中华胃肠外科杂志》2014年第4期383-387,共5页Chinese Journal of Gastrointestinal Surgery

基　　金：国家863计划（2012AA022606）;上海市科学技术委员会资助项目（12401905800）

摘　　要：目的制备荷载氟尿嘧啶（5-Fu）的聚乳酸（PLLA）碳纳米管（CNT）复合材料（5-FU-PLLA-CNTs），并探讨其对人胃癌细胞株（MGC803和MNK45）的体外杀伤效应。方法以PLLA—CNTs为原料，采用超声乳化法荷载5．FU；通过扫描电子显微镜观察5-FU—PLLA—CNTs形态和结构；用紫外可见光光度仪测定不同时间5-FU．PLLA．CNTs的5-FU释放量及累计释放量，并绘制体外释放曲线；设计实验组（不同浓度的5-Fu—PuA—CNTs）及阳性（相应浓度的5-FU）和阴性对照组（不加任何药物），用CCK8法检测不同浓度的5-Fu．PLLA-CNTs对MGC803和MNK45的增殖抑制作用；流式细胞术检测5-FU—PuIA—CNTs作用前后细胞凋亡的变化。结果成功制备5-FU．PLLA．CNTs药物深层薄膜，薄膜载药率为（4．54±0．43）％，包封率（21．56±2．36）％。体外释放实验显示，5-FU-PLLA-CNTs在24h内释放率为23．9％，呈缓慢上升趋势，至31d体外累计释放率达85．3％。CCK8实验显示，与对照组比较，5．Fu—PLLA．CNTs对两株胃癌细胞具有明显抑制作用（P〈0．01），随着药物浓度的增加，抑制作用增强；随着作用时间的延长，呈持续抑制状态。流式细胞仪显示，经1mg／孔5-Fu-PuJA。CNTs处理的实验组MGC803和MNK45细胞凋亡率与阴性对照组比较，差异具有统计学意义（均P〈0．05）；与阳性对照组比较，差异无统计学意义（均P〉0．05）。结论5-FU-PLLA-CNTs具有良好的药物缓释性能，对胃癌细胞株具有明显的杀伤和抑制增殖作用，其最佳浓度为1mg／孔。Objective To prepare cisPLLAtin-loaded polylactic acid/cnts, and to study the anti-tumor effect of 5-FU-PLLA-CNTs on human gastric carcinoma cell lines （MGC803 and MNK45）. Methods 5-FU-PLLA-CNTs were prepared with ultrasound emulsification. The morphology of 5-FU- PLLA-CNTs was determined by scanning electron microscope （SEM）, and its drug loading and drug release curve in vitro were detected by UV-Vis-NIR spectrophotometer. Cells were divided into experiment, positive control and negative control groups. CCK8 method was used to test the cytotoxic effect of 5-FU-PLLA-CNTs in different concentrations on MGC803 and MNK45 cell proliferation. Flow cytometry was employed to measure the apoptotic rate of MGC803 and MNK45 cells before and after the intervention of 5-FU-PLLA-CNTs. Results Deep layer film of 5-FU-PLLA-CNTs was successfully established, whose drug-load rate was （4.54±0.43）%, entrapment rate was （21.56±2.36）%. In vitro release test showed release rate within 24 h of 5-FU-PLLA-CNTs was 23.9% in a aslowly increasing manner, and accumulating release rate was 85.3% at day 31. CCk8 experiment revealed, as compared to control group, 5-FU-PLLA-CNTs significantly inhibited the proliferation of two cell lines in dose- dependent and time-dependent manner. The best 5-FU-PLLA-CNTs concentration of inhibition for human gastric cancer cell lines was 1 mg/well. Flow cytometry indicated the apoptotic rate of MGC803 and MNK45 cells in experiment group treated by 1 rag/well 5-FU-PLLA-CNTs significantly increased as compared to negative control group （P〈0.05）, while the difference was not significant as compared to positive control group （P〉0.05）. Conclusion The 5-FU-PLLA-CNTs has good drug sustained-release capacity, and can significantly kill and inhibit the proliferation of MGC803 and MNK45 cell lines.

关 键 词：胃肿瘤 氟尿嘧啶 聚乳酸 体外杀伤效应 

分 类 号：R735.2[医药卫生—肿瘤]