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

Yamamoto, Kumiko; Ikenaka, Yoshinori; Ichise, Takahiro; Bo, Tomoki; Ishizuka, Mayumi; Yasui, Hironobu; Hiraoka, Wakako; Yamamori, Tohru; Inanami, Osamu

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

Yamamoto, Kumiko; Ikenaka, Yoshinori; Ichise, Takahiro et al.

Free Radical Research, 2018, 52(6), 648-660

Permalink : https://hdl.handle.net/2115/73690

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アクセス権	open access
URI	https://hdl.handle.net/2115/73690
タイトル	Evaluation of mitochondrial redox status and energy metabolism of X-irradiated HeLa cells by LC/UV, LC/MS/MS and ESR
著者	Yamamoto, Kumiko ; Ikenaka, Yoshinori ; Ichise, Takahiro ; Bo, Tomoki ; Ishizuka, Mayumi ; Yasui, Hironobu ; Hiraoka, Wakako ; Yamamori, Tohru ; Inanami, Osamu
著者名 Yamamoto, Kumiko
e-Rad_Researcher 40543509 著者名 Ikenaka, Yoshinori
著者名 Ichise, Takahiro
著者名 Bo, Tomoki
e-Rad_Researcher 50332474 著者名 Ishizuka, Mayumi
e-Rad_Researcher 10570228 著者名 Yasui, Hironobu
著者名 Hiraoka, Wakako
e-Rad_Researcher 00512675 著者名 Yamamori, Tohru
e-Rad_Researcher 10193559 著者名 Inanami, Osamu
言語	英語
キーワード	tumour
	ESR
	Fe-S cluster
	ionizing radiation
	mitochondrial ETC
	semiquinone
	tumour
発行日	2018-04-19
出版者	Taylor & Francis
収録物名	Free Radical Research
巻(号)	52(6)
ページ	648-660
抄録	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.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	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 .
PISSN	1071-5762
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1080/10715762.2018.1460472