博宁霉素克服人肿瘤细胞多药耐药性的机制  

Mechanism overcoming multidrug resistance of human tumor cells by boningmycin

作　　者：张秀敏[1] 许鸿章[1] 鲁敏[1] 陈汝贤[1] 何琪杨[1] 

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所,北京100050

出　　处：《中国新药杂志》2012年第12期1332-1336,1340,共6页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项(2012ZX09301002-001)

摘　　要：目的:研究博来霉素族新抗生素博宁霉素能否克服肿瘤细胞的多药耐药性。方法:选用耐多柔比星的人乳腺癌MCF-7细胞(MCF-7/DOX)、耐长春新碱的人口腔上皮癌KB细胞(KBV200)及其相应的敏感细胞。MTT法检测细胞增殖的抑制作用;流式细胞仪检测细胞周期的变化以及细胞内活性氧自由基的水平;DNA特异染料Hoechst 33342染色检测染色质凝集;Western blot检测蛋白的表达。结果:MTT法检测到博宁霉素抑制MCF-7/DOX和敏感MCF-7细胞的IC50值分别为0.17和0.10μmol·L-1;抑制KBV200和敏感细胞的IC50值分别为0.38和0.21μmol·L-1。耐药细胞对博宁霉素没有交叉耐药性。博宁霉素使耐药细胞停滞在G2/M期,细胞内活性氧自由基水平增加,染色质发生凝集,引起细胞凋亡,Western blotting检测到降低多药耐药相关蛋白P-糖蛋白的含量。结论:博宁霉素能克服肿瘤细胞的多药耐药性,其作用机制与引起细胞凋亡和降低P-糖蛋白表达有关。Objective： To investigate whether boningmycin,a new member of bleomycin family,overcomes multidrug resistance of tumor cells.Methods： The doxorubicin-resistant human breast cancer MCF-7 cells（MCF-7/DOX）and vincristine-resistant human oral epithelial cells（KBV200）and related sensitive cells were chosen for the experiments.MTT reduction assay was used to detect the inhibitory effect on cell proliferation.Cell cycle distribution and accumulation of intracellular reactive oxygen species（ROS） were analyzed with flow cytometry.Chromatin condensation was detected by a fluorescent microscope after Hoechst 33342 staining.Protein expression was detected by Western blot.Results： MTT results showed that IC50 values of MCF-7/DOX cells and MCF-7 cells inhibited by boningmycin were 0.17 and 0.10 μmol·L-1,respectively,and those of KBV200 cells and KB cells were 0.38 and 0.21 μmol·L-1,respectively,showing no cross resistance to boningmycin in resistant cells.Furthermore,the arrest at G2/M phase and increase in ROS level were detected by flow cytometry.Boningmycin induced the chromatin condensation,apoptosis and decreased expression of multidrug resistance-associated P-glycoprotein detected by Western blotting.Conclusion： Boningmycin can overcome the multidrug resistance in tumor cells and its mechanisms are related with induction of apoptosis and reduction of P-glycoprotein expression.

关 键 词：博宁霉素 多药耐药性 P-糖蛋白 细胞凋亡 博来霉素类 

分 类 号：R979.1[医药卫生—药品]