A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis
A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis
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DOI：
10.1002/cyto.b.20004
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发表时间：
2004-05-01
影响因子：
3.4
通讯作者：
Boutonnat, J
中科院分区：
文献类型：
--
作者：
Barbier, M;Gray, BD;Boutonnat, J
背景：肿瘤细胞的增殖状态和多药耐药在决定细胞对化疗的反应中起着关键作用。方法：采用Hoechst 33342（DNA）、JC 1（DNA）、JC 1（DNA）和DNA三色标记技术，对肿瘤细胞进行标记，并对标记后的细胞进行形态学观察，同时观察肿瘤细胞的多药耐药（MDR）表型、增殖状态、凋亡相关的线粒体电位变化。（线粒体电位）和远红荧光膜嵌入染料：PTIR 271结果：DNA和线粒体电位的联合染色允许鉴定表达P-糖蛋白（Pgp）的亚群和由Pgp介导的MDR表型，在Pgp阴性亚群中，鉴定凋亡细胞并评价活细胞的细胞周期状态。此外，远红荧光膜嵌入染料，PTIR 271，允许同时监测细胞分裂状态的染料稀释在两个药物敏感和耐药population.Conclusion：这种三重标记是一个有趣的方法来研究药物敏感性和耐药性的增殖状态在活的肿瘤细胞。(C)2004 Wiley-Liss，Inc.
Background: Proliferative status and multidrug resistance play a key role in determining cell response to chemotherapy. There is a need to develop multiple labeling method that allows simultaneous assessment of multidrug resistant (MDR) phenotype, proliferative status, apoptosis related changes in mitochondrial potential, in chemosensitive and chemoresistant tumor cell populations.Methods: A three-color labeling was performed using Hoechst 33342 (DNA), JC1 (mitochondrial potential), and a far red fluorescent membrane intercalating dye: PTIR271 (proliferation).Results: Combined staining of DNA and mitochondrial potential allows identification of subpopulations expressing and MDR phenotype mediated by P-glycoprotein (Pgp), and, in Pgp negative subpopulations, identification of apoptotic cells and evaluation of cell cycle status in viable cells. Addition of a far red fluorescent membrane intercalating dye, PTIR271, allows simultaneous monitoring of cell division status by dye dilution in both drug sensitive and drug resistant populations.Conclusion: This triple labeling is an interesting method to study the proliferation status of drug sensitivity and drug resistance in viable tumor cells. (C) 2004 Wiley-Liss, Inc.