期刊
ACS SENSORS
卷 4, 期 6, 页码 1619-1625出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.9b00344
关键词
double-mediator system; large-scale integration-based amperometric device; NAD(P)H:quinone oxidoreductase 1; cell analysis; cell aggregates
资金
- Japan Society for the Promotion of Science (JSPS) [16H02280, 18H01840, 18H01999]
- Program for Creation of Interdisciplinary Research from Frontier Research Institute for Interdisciplinary Sciences, Tohoku University
- Grants-in-Aid for Scientific Research [18H01840, 18H01999] Funding Source: KAKEN
NAD(P)H:quinone oxidoreductase 1 (NQO1) is a key enzyme providing cytoprotection from quinone species. In addition, it is expressed at high levels in many human tumors, such as breast cancer. Therefore, it is considered to be a potential target in cancer treatment. In order to detect intracellular NQO1 activity in MCF-7 aggregates as a cancer model, we present, in this study, a double-mediator system combined with large-scale integration (LSI)-based amperometric devices. This LSI device contained 20 x 20 Pt working electrodes with a 250 mu m pitch for electrochemical imaging. In the detection system, menadione (MD) and [Fe(CN)(6)](3-) were used. Since MD can diffuse into cells due to its hydrophobicity, it is reduced into menadiol by intracellular NQO1. The menadiol diffuses out of the cells and reduces [Fe(CN)(6)](3-) of a hydrophilic mediator into [Fe(CN)(6)](4-). The accumulated [Fe(CN)(6)](4-) outside the cells is electrochemically detected at 0.5 V in the LSI device. Using this strategy, the intracellular NQO1 activity of MCF-7 aggregates was successfully detected. The effect of rotenone, which is an inhibitor for Complex I, on NQO1 activity was also investigated. In addition, NQO1 and respiration activities were simultaneously imaged using the detection system that was further combined with electrochemicolor imaging. Thus, the double-mediator system was proven to be useful for evaluating intracellular redox activity of cell aggregates.
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