活性氧介导的线粒体功能不全与小细胞肺癌多药耐药细胞H446/CDDP对顺铂耐药性关系的研究  

Study of reactive oxygen species mediated mitochondrial dysfunction on cisplatin resistance in multidrug resistance cells H446 / CDDP

作　　者：王瑞璇[1] 马李杰[1] 段鸿涛 王力彬[3] 金发光[1] 

机构地区：[1]第四军医大学唐都医院呼吸内科,西安710038 [2]第四军医大学唐都医院胸外科,西安710038 [3]第四军医大学药学系,西安710038

出　　处：《中华肺部疾病杂志（电子版）》2015年第5期29-32,共4页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：国家自然科学基金(81071933)

摘　　要：目的探讨人小细胞肺癌多药耐药细胞H446/CDDP细胞内活性氧(ROS)介导的线粒体功能不全及其与顺铂耐药性之间的关系。方法用MTT法检测H446细胞及其多药耐药细胞H446/CDDP在5μg/ml顺铂作用后的细胞生存率,流式细胞术检测细胞凋亡,荧光显微镜观察顺铂诱导后两种细胞内ROS含量以及线粒体膜电位(MMP)水平。实验中均设置空白对照组。结果顺铂诱导后细胞的生存率均随药物作用时间的延长明显降低,且H446/CDDP细胞在各个时间点的生存率均明显高于H446细胞;5μg/ml顺铂作用24 h后H446和H446/CDDP两种细胞的早期凋亡百分比分别为(8.63±0.45)%和(1.97±0.42)%,具有明显的统计学差异(P<0.05);H446细胞内ROS的水平在顺铂诱导24 h后明显升高,而H446/CDDP细胞内ROS的水平无明显变化;与此同时,H446细胞MMP下降水平明显高于H446/CDDP细胞。结论顺铂可使小细胞肺癌H446细胞内ROS的产生增加,MMP下降,从而诱导细胞凋亡,而多药耐药细胞H446/CDDP顺铂耐药性的产生与顺铂作用后细胞内ROS减少,MMP下降抑制所致的线粒体介导的内源性细胞凋亡途径耐受有关。Objective To explore the relationship between reactive oxygen species-dependent mitochondrial dysfunction and cisplatin resistance in multi-drug resistance cells H446 / CDDP. Methods The survival rate of small cell lung cancer cells H446 and the multi-drug resistance H446 / CDDP cells were measured by MTT assay after cisplatin induced（ 5 μg / ml）. Cell apoptosis were detected by flow cytometry.Meanwhile both Intracellular reactive oxygen species（ ROS） and mitochondrial membrane potential（ MMP）were observed using fluorescence microscope. There were blank controls in the study. Result The survival rate of both cells were obviously decreased after cisplatin induced as stimulating time prolonged,and at each time point H446 / CDDP cells survival rate were significantly higher than that of H446 cells. Early apoptosis percentage of H446 and H446 / CDDP after cisplatin induced（ 5 μg / ml） were（ 8. 63 ± 0. 45） % and（ 1. 97 ±0. 42） %,the difference is statistically significant（ P 〈 0. 05）. ROS content in H446 cells increased markly after being stimulated 24 h by cisplatin,on the other hand ROS in H446 / CDDP cells barely rose. At the same time,reduction of the H446 cells MMP was significantly higher than that of H446 / CDDP cells. Conclusion Cisplatin induced apoptosis involves increased production of ROS,and dissipation of MMP,while cisplatin resistance in multi-drug resistance cells H446 / CDDP is associated with resistance of mitochondriamediated apoptotic（ intrinsic） pathways which is mediated by the reduction of intracellular ROS and the inhibition of loss of MMP after the application of cisplatin.

关 键 词：小细胞肺癌 H446/CDDP细胞 顺铂 细胞凋亡 活性氧 

分 类 号：R737.14[医药卫生—肿瘤]