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Evaluation of mitochondrial redox status and energy metabolism of X-irradiated HeLa cells by LC/UV, LC/MS/MS and ESR

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Title: Evaluation of mitochondrial redox status and energy metabolism of X-irradiated HeLa cells by LC/UV, LC/MS/MS and ESR
Authors: Yamamoto, Kumiko Browse this author
Ikenaka, Yoshinori Browse this author →KAKEN DB
Ichise, Takahiro Browse this author
Bo, Tomoki Browse this author
Ishizuka, Mayumi Browse this author →KAKEN DB
Yasui, Hironobu Browse this author →KAKEN DB
Hiraoka, Wakako Browse this author
Yamamori, Tohru Browse this author →KAKEN DB
Inanami, Osamu Browse this author →KAKEN DB
Keywords: tumour
ESR
Fe-S cluster
ionizing radiation
mitochondrial ETC
semiquinone
tumour
Issue Date: 19-Apr-2018
Publisher: Taylor & Francis
Journal Title: Free Radical Research
Volume: 52
Issue: 6
Start Page: 648
End Page: 660
Publisher DOI: 10.1080/10715762.2018.1460472
Abstract: To evaluate the metabolic responses in tumour cells exposed to ionizing radiation, oxygen consumption rate (OCR), cellular lipid peroxidation, cellular energy status (intracellular nucleotide pool and ATP production), and mitochondrial reactive oxygen species (ROS), semiquinone (SQ), and iron-sulphur (Fe-S) cluster levels were evaluated in human cervical carcinoma HeLa cells at 12 and 24h after X-irradiation. LC/MS/MS analysis showed that levels of 8-iso PGF(2 alpha) and 5-iPF(2 alpha)VI, lipid peroxidation products of membrane arachidonic acids, were not altered significantly in X-irradiated cells, although mitochondrial ROS levels and OCR significantly increased in the cells at 24h after irradiation. LC/UV analysis revealed that intracellular AMP, ADP, and ATP levels increased significantly after X-irradiation, but adenylate energy charge (adenylate energy charge (AEC) = [ATP + 0.5 x ADP]/[ATP +ADP + AMP]) remained unchanged after X-irradiation. In lowtemperature electron spin resonance (ESR) spectra of HeLa cells, the presence of mitochondrial SQ at g = 2.004 and Fe-S cluster at g = 1.941 was observed and X-irradiation enhanced the signal intensity of SQ but not of the Fe-S cluster. Furthermore, this radiation-induced increase in SQ signal intensity disappeared on treatment with rotenone, which inhibits electron transfer from Fe-S cluster to SQ in complex I. From these results, it was suggested that an increase in OCR and imbalance in SQ and Fe-S cluster levels, which play a critical role in the mitochondrial electron transport chain (ETC), o(cur after X-irradiation, resulting in an increase in ATP production and ROS leakage from the activated mitochondrial ETC.
Rights: This is an Accepted Manuscript of an article published by Taylor & Francis in Free Radical Research in 2018, available online: http://www.tandfonline.com/10.1080/10715762.2018.1460472 .
Type: article (author version)
URI: http://hdl.handle.net/2115/73690
Appears in Collections:獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 稲波 修

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