Combination of Double-Mediator System with Large-Scale Integration-Based Amperometric Devices for Detecting NAD(P)H:quinone Oxidoreductase 1 Activity of Cancer Cell Aggregates

Kosuke Ino, Takehiro Onodera, Mika T. Fukuda, Yuji Nashimoto, Hitoshi Shiku

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

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 × 20 Pt working electrodes with a 250 μm 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.

Original languageEnglish
Pages (from-to)1619-1625
Number of pages7
JournalACS Sensors
Volume4
Issue number6
DOIs
Publication statusPublished - 2019 Jun 28

Keywords

  • NAD(P)H:quinone oxidoreductase 1
  • cell aggregates
  • cell analysis
  • double-mediator system
  • large-scale integration-based amperometric device

ASJC Scopus subject areas

  • Bioengineering
  • Instrumentation
  • Process Chemistry and Technology
  • Fluid Flow and Transfer Processes

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