流式细胞技术分析多壁碳纳米管对人多药耐药性胶质瘤细胞活力的影响  

Flow Cytometric Analysis of Multi-Walled Carbon Nanotubes' Effect on Human Glioma Cell Viability

作　　者：徐永红[1] 邢怡桥[1] 陈效[2] 

机构地区：[1]武汉大学人民医院眼科,湖北武汉430060 [2]华中科技大学同济药学院,湖北武汉430030

出　　处：《武汉大学学报（医学版）》2011年第6期727-732,共6页Medical Journal of Wuhan University

摘　　要：目的:建立荧光探针染色-流式细胞检测术(FCM)的方法用于分析多壁碳纳米管(MWCNTs)对人多药耐药性(MDR)胶质瘤细胞活力的影响。方法:将人MDR胶质瘤细胞给予不同浓度的MWCNTs处理24h。随后以一系列反映细胞功能不同方面的荧光探针(JC-1、R123、FDA和PI)在原位(in situ)和非原位(ex situ)两种条件下对细胞进行染色,以FCM对各个探针的细胞染色情况进行检测。结果:在经过MWCNTs处理的细胞中,R123染色在in situ条件下呈浓度依赖性增强,但在ex situ条件下无明显改变。JC-1染色在in situ条件下呈显著浓度依赖性降低,但在ex situ条件下仅略微减弱。FDA染色在in situ条件下随MWCNTs浓度增加呈"钟形"改变,而在ex situ条件呈小幅度增强。PI染色阳性的细胞数在两种条件下均略微增加。结论:荧光探针染色结合FCM在经过验证优化以后可有效地用于研究MWCNTs的细胞效应。在本实验的MWCNTs处理浓度和时间条件下,人MDR胶质瘤细胞的总体活力未受明显影响。但涉及细胞膜的一些功能可能发生改变。Objective： To establish and optimize a series of methods based on fluorescent probe staining in combination with flow cytometry（FCM） to investigate the effect of multi-walled carbon nanotubes（MWCNTs） on cell viability.Methods： The human multi-drug resistance（MDR） glioma cells were exposed to MWCNTs for 24 h and subsequently stained with a panel of fluorescent probes including JC-1,R123,FDA,and PI.Staining was performed in situ and ex situ and was analyzed by FCM.Results： In MWCNT-exposed cells,R123 staining was significantly enhanced under in situ condition but varied little from control under ex situ condition.Significant decrease of JC-1 staining was observed in in situ cells but not in ex situ cells.FDA staining displayed a somewhat bell-shaped pattern in in situ cells but showed minor enhancement in ex situ cells.The number of cells positive of PI staining only slightly increased under both conditions.Conclusion： Fluorescent probe staining combined with FCM is a convenient and effective approach for investigating cellular responses to MWCNTs exposure.In the current work,human MDR glioma cell viability was generally maintained in the presence of MWCNTs.But certain cellular functions associated with cell membrane were probably affected.

关 键 词：多壁碳纳米管:胶质瘤细胞 流式细胞检测 细胞活力 

分 类 号：R73-3[医药卫生—肿瘤]