摘要
目的研究5氟尿嘧啶半乳糖神经酰胺脂质体(5FuGCL)体外对耐5FuHepG2细胞株的抑制作用并对其抗耐药机制进行探讨。方法在建立耐5Fu的HepG2细胞株模型的基础上,采用MTT法检测5FuGCL对其的抑制作用。另外,通过高效液相法(HPLC)检测细胞内液的药物含量、免疫组化法检测胸苷酸合酶(TS)的表达、化学法检测NO含量等研究其抗耐药作用机制。结果5FuGCL(75,150,300,600,1200μmol·L-1)对耐5Fu的HepG2细胞株有明显的抑制作用,IC50为158.6μmol·L-1,远小于游离5Fu(400.9μmol·L-1);5FuGCL(300μmol·L-1)对耐5Fu的HepG2细胞的抑制作用随时间的延长而增强,其中(24～48h)的抑制作用明显强于相同浓度的游离5Fu。5FuGCL(300μmol·L-1)与相同浓度的游离5Fu比较,能明显增加药物进入HepG2细胞内液的程度;5FuGCL(75,300,1200μmol·L-1)既可明显降低TS的表达,又可显著增加NO的含量,且5FuGCL(300,1200μmol·L-1)与相同浓度的游离5Fu相比,其作用明显增强。结论5FuGCL有明显的抗5Fu耐药作用,这可能是通过增加细胞内液5Fu的含量,抑制TS的表达和增加NO含量实现的。
Aim To study inhibitory effect of 5-fluorouracil encapsulated by galactosylceramide liposomes （5- Fu-GCL）on 5-Fu-resistent HepG2 cells and its mechanisms. Methods Inhibitory effect of 5-Fu-GCL on established model of 5-Fu-resistant HepG2 cells was assessed with MTT assay in vitro. The concentration-time course of 5-Fu-GCL in intracellular fluid was detected with high performance liquid chromatography （HPLC）. Thymidylic acid synthase （ TS ） expression was observed with immunohistochemical method, and NO content was determined with chemical method. Resuits Obvious inhibitory effects of 5-Fu-GCL （75, 150,300,600, 1200μmol· L^-l ） on 5-Fu-resistant HepG2 cells were observed with IC50 of 158.6μmol· L^-l, far lower than that of free 5-Fu （400. 9 μmol·L^-1）. 5-Fu-GCL （300μmol·L^-1） inhibited 5-Fu-resistant HepG2 cells in a time-dependent manner, and the inhibitory effect of 5-Fu-GCL was stronger than that of free 5-Fu during 12～48 h. Compared with free 5- Fu,5-Fu-GCL （300μmol·L^-1 ） increased the content of intracellular fluid in 5-Fu-resistant HepG2 cells. 5-Fu-GCL（62.5,300,1200μmol·L^-1） not only inhibited the expression of TS, but also increased the production of NO in 5-Fu-resistant HepG2 cells,and these effects of 5-Fu-GCL （300, 1200 μmol· L^-1 ） were stronger than those of free 5-Fu. Conclusion 5-Fu- GCL has inhibitory effect on 5-Fu-resistant HepG2 cells. The effect may be related to the increased concentration of 5-Fu-GCL in intracellular fluid, inhibited expression of TS and increased production of NO.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2005年第8期954-958,共5页
Chinese Pharmacological Bulletin
基金
国家教育部青年骨干教师基金资助项目(No1869)
安徽省教育厅青年教师基金资助项目(No2000jp112)
关键词
5-氟尿嘧啶
半乳糖神经酰胺
脂质体
耐药
5-fluorouracil （5-Fu）
galactosylceramide （ GC ）
liposome
drug resistance
