摘要
目的在同一实验系统中比较Ru-486、MPA、VP、CsA对非P-g介导的卵巢癌耐顺铂细胞株COC1/DDP耐药性的逆转作用并对其作用机制进行探讨,为临床克服耐药提供参考依据。方法采用MTT法从细胞代谢,DNA凝胶电泳从细胞凋亡方面系统观察比较DDP、Ru-486、MPA、VP、CsA 5种药物对卵巢癌多药耐药细胞系COC1/DDP的影响及相互关系,检测上述5种药物与COC1/DDP作用时细胞内GSH-ST和GSH-Px活性的变化,并采用westernblotting以Bcl-2抗体为分子探针对上述5种药物与COC1/DDP作用时细胞内Bcl-2基因的表达进行测定对其逆转机制进行探讨。结果1)Ru-486、CsA、MPA对COC1/DDP有直接抑制作用,其抑制率随浓度增高进行性升高,而VP对COC1/DDP没有直接抑制作用。和DDP联用时,Ru-486、MPA、VP、CsA都能显著增强COC1/DDP对DDP的敏感性,达到一定浓度后,可以完全逆转COC1/DDP对DDP的耐药性。这种逆转是通过抑制细胞代谢、诱导细胞凋亡的结果。2)单纯2.5μg/ml的DDP能使GSH-Px及GST活力上升,除VP外,Ru-486等药物可降低GSH-Px和GST活力,与DDP联用时亦如此。提示,Ru-486、CsA、MPA可以通过抑制GSH转移酶系统阻止对细胞损伤的修复而达到对耐药性的逆转。3)单独的DDP、Ru--486等药物非但对Bcl-2的表达没有抑制作用,反而促进了其表达,以MPA最甚。与DDP联用时,却出现了一个有趣的现象,即可以抑制Bcl-2的表达,Ru-486与DDP联用时Bcl-2的表达几乎完全被阻截。结论:VP、Ru-486、CsA和MPA对卵巢癌多药耐药细胞系COC1/DDP的耐药性有确切逆转作用。其逆转机制可能与降低GSH转移酶系统活性有关。当与DDP联用时可以通过降低Bcl-2蛋白表达来逆转耐药。
Objective Ovarian malignant tumor is one of the three most harmful female genital tumor. For lack of efficient treatment,the five-year-survival-rate have long been lowered at the range of 25%-30%. Multi-drug resistance （MDR）is the major reason that results the patient a bad consequence when chemical therapy was done. To investigation the MDR inhibitor has become a very important mission of the tumor chemical therapy since verapamil was found that could be used as a MDR reverser in 1981.Our study is aim to observe systemically the influence of five drugs（DDP, Ru-486,MPA,Vp,CsA）on the non-P-glycoprotein-translated cisplatin resistance cell lines COC1/DDP and their mutual relationships,and try to investigate the mechanism of the reversion. Methods Using MTT to observe the effect of reversion. Apoptosis DNA gel electrophoresis,GSH-ST and GSH-Px analysis as well as western blotting were used to approach the mechanism of reversion. Results Ru-486,CsA,MPA can repress COC1/DDP directly,their repressive rate is in exact ratio to their concentration. VP has not this direct repressive ability. Combined with DDP,all of Ru-486,CsA, MPA,VP can enhance the susceptibility of DDP,when certain concentration is reached,the resistance of COC1/DDP can be totally reversed. This reversion is caused by the metabolism repression,apoptosis induction and DNA damage of cells. Use 2.5 μg/ml DDP can made GSH-Px and GST more active. Except VP,the other three drugs can reduce the activity of GSH-Px and GST,and the result is the same when combined with DDP. It suggest that Ru-486,CsA,MPA can reverse the MDR through repress the GSH transferase system which can repair the damage of cells. Use single drug, such as DDP,Ru-486 et al,can not repress the expression of Bcl-2,they promote the expression,especially when MPA was singly used. But when the drugs were used combined with DDP,an interest phenomenon appeared,they can reduce the expression of Bcl-2 gene. The expression of Bcl-2 was wholly inhabited when Ru-486 was used with the combination of DDP. Conclusion Ru-486,CsA,MPA and VP all definitely have reverse effect on the MDR of COC1/DDP. The mechanism of the reversion is probably the drugs could repress the GSH transferase system which can repair the damage of cells and reduce the expression of Bcl-2 while the drugs used combined with DDP.
出处
《生物技术通报》
CAS
CSCD
2008年第2期172-179,共8页
Biotechnology Bulletin
基金
国家自然科学基金资助项目(30371619)
