Real-time cell viability assays using a new anthracycline derivative DRAQ7®.
Real-time cell viability assays using a new anthracycline derivative DRAQ7®.
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DOI：
10.1002/cyto.a.22228
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发表时间：
2013-02
期刊：
影响因子：
3.7
通讯作者：
Wlodkowic, Donald
中科院分区：
文献类型：
--
作者：
Akagi, Jin;Kordon, Magdalena;Zhao, Hong;Matuszek, Anna;Dobrucki, Jurek;Errington, Rachel;Smith, Paul J.;Takeda, Kazuo;Darzynkiewicz, Zbigniew;Wlodkowic, Donald
关键词：
被完整的细胞排除带电的荧光染料已经成为一种确定细胞活力的成熟的方法。为了寻找一种能够长期实时监测细胞死亡的非侵入性荧光探针，我们评估了一种新的蒽环类化合物DRAQ7。这种新的探针不会穿透活细胞的质膜，但当细胞膜的完整性受到损害时，它会进入并很容易地与核DNA结合，报告细胞死亡。事实证明，它对一组连续生长长达72小时的癌细胞株无毒，在使用激光扫描显微镜和流式细胞仪进行分析时，也不会引发任何可检测到的DNA损伤信号。DRAQ7提供了一种灵敏的、实时的读出细胞死亡的方法，这些应激源包括低氧、饥饿和药物诱导的细胞毒性。在DRAQ7存在的情况下，对抗癌药的总体反应和由此产生的药理剂量-反应曲线不受肿瘤细胞生长的影响。此外，我们首次介绍了一种基于DRAQ7和线粒体内膜电位(ΔΨm)敏感探针TMRM相结合的近实时微量流式细胞仪分析方法。我们提供的证据表明，这种低剂量、实时标记的方法提供了抗癌药物作用的多参数和动力学指纹图谱。
The exclusion of charged fluorescent dyes by intact cells has become a well-established assay for determining viability of cells. In search for a non-invasive fluorescent probe capable of long-term monitoring of cell death in real-time, we evaluated a new anthracycline derivative DRAQ7. The novel probe does not penetrate the plasma membrane of living cells but when the membrane integrity is compromised, it enters and binds readily to nuclear DNA to report cell death. It proved to be non-toxic to a panel of cancer cell lines grown continuously for up to 72 hours and did not induce any detectable DNA damage signaling when analyzed using laser scanning microscopy and flow cytometry. DRAQ7 provided a sensitive, real-time readout of cell death induced by a variety of stressors such as hypoxia, starvation and drug-induced cytotoxicity. The overall responses to anti-cancer agents and resulting pharmacological dose-response profiles were not affected by the growth of tumor cells in the presence DRAQ7. Moreover, we for the first time introduced a near real-time microflow cytometric assay based on combination of DRAQ7 and mitochondrial inner membrane potential (ΔΨm) sensitive probe TMRM. We provide evidence that this low-dosage, real-time labeling procedure provides multi-parameter and kinetic fingerprint of anti-cancer drug action.
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DOI：
10.4161/cc.9.12.11911
发表时间：
2010-06-15
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Skommer J;Darzynkiewicz Z;Wlodkowic D
通讯作者：
Wlodkowic D
影响因子：
3.7
作者：
Zhao, Hong;Halicka, H. Dorota;Traganos, Frank;Jorgensen, Ellen;Darzynkiewicz, Zbigniew
通讯作者：
Darzynkiewicz, Zbigniew
影响因子：
4
作者：
Floryk, D;Houstek, J
通讯作者：
Houstek, J
影响因子：
7.4
作者：
Khoshmanesh, Khashayar;Akagi, Jin;Wlodkowic, Donald
通讯作者：
Wlodkowic, Donald
影响因子：
7.4
作者：
Khoshmanesh, Khashayar;Akagi, Jin;Wlodkowic, Donald
通讯作者：
Wlodkowic, Donald