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
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-PLLA-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-PLLA-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24 h内释放率为23.9%,呈缓慢上升趋势,至31 d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1 mg/孔5-FU-PLLA-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1 mg/孔。
更多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 mg/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.
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