Low-Temperature EPR Spectroscopy as a Probe-Free Technique for Monitoring Oxidants Formed in Tumor Cells and Tissues: Implications in Drug Resistance and OXPHOS-Targeted Therapies.
Low-Temperature EPR Spectroscopy as a Probe-Free Technique for Monitoring Oxidants Formed in Tumor Cells and Tissues: Implications in Drug Resistance and OXPHOS-Targeted Therapies.
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DOI：
10.1007/s12013-018-0858-1
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发表时间：
2019-03
影响因子：
2.6
通讯作者：
Bennett B
中科院分区：
文献类型：
--
作者：
Kalyanaraman B;Cheng G;Zielonka J;Bennett B
由氧化和硝化代谢形成的氧化剂包括活性氧物种(ROS)，如超氧化物、过氧化氢/脂质过氧化氢和活性氮物种(RNS)(例如过氧亚硝酸盐[ONOO-]和二氧化氮)，以及活性卤化物种(次氯酸[HOCl])。ROS和RNS越来越多地参与肿瘤的发生以及肿瘤的生长、进展和转移。最近，ROS被认为与免疫治疗中的耐药、代谢重编程和T细胞代谢有关。大多数情况下，荧光探针已用于细胞培养系统。通过对诊断标记产物的LC-MS分析，获得了物种的同一性。然而，如果不是不可能的话，将这些化验方法外推到癌症移植瘤是困难的。因此，发展一种无探针的方法来监测和评估肿瘤细胞和肿瘤移植瘤中氧化剂的形成是至关重要的和及时的。在这里，我们描述了低温下体外电子顺磁共振(EPR)光谱的使用，作为一种独特的无探针技术，通过来自线粒体和胞质氧化还原蛋白质中氧化还原中心的EPR信号，特别是铁-硫簇，来评估细胞内的氧化和氧化剂。肿瘤细胞受到线粒体氧化磷酸化抑制的例子被提出。这种体外方法可以很容易地扩展到监测从小鼠和人类分离的肿瘤组织中氧化剂的形成。
Oxidants formed from oxidative and nitrative metabolism include reactive oxygen species (ROS) such as superoxide, hydrogen peroxide/lipid hydroperoxides and reactive nitrogen species (RNS) (e.g., peroxynitrite [ONOO–] and nitrogen dioxide), and reactive halogenated species (hypochlorous acid [HOCl]). Increasingly, ROS and RNS are implicated in tumorigenesis as well as tumor growth, progression, and metastasis. Recently, ROS were implicated in drug resistance, metabolic reprogramming, and T-cell metabolism in immunotherapy. Mostly, fluorescent probes have been used in cell culture systems. The identity of species is obtained by LC–MS analyses of diagnostic marker products. However, extrapolation of these assays to cancer xenografts is difficult if not impossible. Thus, development of a probe-free assay for monitoring and assessing oxidant formation in tumor cells and tumor xenografts is critical and timely. Here, we describe the use of ex vivo electron paramagnetic resonance (EPR) spectroscopy at cryogenic temperatures as a uniquely useful probe-free technique for assessing intracellular oxidation and oxidants via EPR signals from redox centers, particularly iron-sulfur clusters, in mitochondrial and cytosolic redox proteins. Examples of cancer cells subjected to inhibition of mitochondrial oxidative phosphorylation are presented. This ex vivo methodology can be readily extended to monitor oxidant formation in tumor tissues isolated from mice and humans.
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影响因子：
2.9
作者：
Bulteau, AL;Ikeda-Saito, M;Szweda, LI
通讯作者：
Szweda, LI
DOI：
10.1073/pnas.1620433114
发表时间：
2017-01-31
影响因子：
11.1
作者：
Chamoto, Kenji;Chowdhury, Partha S.;Honjo, Tasuku
通讯作者：
Honjo, Tasuku
影响因子：
4.1
作者：
Dunne, Jacqueline;Caron, Alexis;Cooper, Chris E.
通讯作者：
Cooper, Chris E.
DOI：
10.1111/j.1432-1033.1975.tb04128.x
发表时间：
1975-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
BEINERT, H;ACKRELL, BAC;SINGER, TP
通讯作者：
SINGER, TP
二甲双胍的线粒体靶向类似物在胰腺癌细胞中表现出增强的抗增殖和放射增敏作用。
影响因子：
11.2
作者：
Cheng G;Zielonka J;Ouari O;Lopez M;McAllister D;Boyle K;Barrios CS;Weber JJ;Johnson BD;Hardy M;Dwinell MB;Kalyanaraman B
通讯作者：
Kalyanaraman B