Mitochondrial respiratory function induces endogenous hypoxia.

Hypoxia influences many key biological functions. In cancer, it is generally believed that hypoxic condition is generated deep inside the tumor because of the lack of oxygen supply. However, consumption of oxygen by cancer should be one of the key means of regulating oxygen concentration to induce h...

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Main Authors: Sara Prior, Ara Kim, Toshitada Yoshihara, Seiji Tobita, Toshiyuki Takeuchi, Masahiro Higuchi
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0088911&type=printable
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author Sara Prior
Ara Kim
Toshitada Yoshihara
Seiji Tobita
Toshiyuki Takeuchi
Masahiro Higuchi
author_facet Sara Prior
Ara Kim
Toshitada Yoshihara
Seiji Tobita
Toshiyuki Takeuchi
Masahiro Higuchi
author_sort Sara Prior
collection DOAJ
description Hypoxia influences many key biological functions. In cancer, it is generally believed that hypoxic condition is generated deep inside the tumor because of the lack of oxygen supply. However, consumption of oxygen by cancer should be one of the key means of regulating oxygen concentration to induce hypoxia but has not been well studied. Here, we provide direct evidence of the mitochondrial role in the induction of intracellular hypoxia. We used Acetylacetonatobis [2-(2'-benzothienyl) pyridinato-kN, kC3'] iridium (III) (BTP), a novel oxygen sensor, to detect intracellular hypoxia in living cells via microscopy. The well-differentiated cancer cell lines, LNCaP and MCF-7, showed intracellular hypoxia without exogenous hypoxia in an open environment. This may be caused by high oxygen consumption, low oxygen diffusion in water, and low oxygen incorporation to the cells. In contrast, the poorly-differentiated cancer cell lines: PC-3 and MDAMB231 exhibited intracellular normoxia by low oxygen consumption. The specific complex I inhibitor, rotenone, and the reduction of mitochondrial DNA (mtDNA) content reduced intracellular hypoxia, indicating that intracellular oxygen concentration is regulated by the consumption of oxygen by mitochondria. HIF-1α was activated in endogenously hypoxic LNCaP and the activation was dependent on mitochondrial respiratory function. Intracellular hypoxic status is regulated by glucose by parabolic dose response. The low concentration of glucose (0.045 mg/ml) induced strongest intracellular hypoxia possibly because of the Crabtree effect. Addition of FCS to the media induced intracellular hypoxia in LNCaP, and this effect was partially mimicked by an androgen analog, R1881, and inhibited by the anti-androgen, flutamide. These results indicate that mitochondrial respiratory function determines intracellular hypoxic status and may regulate oxygen-dependent biological functions.
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spelling doaj-art-c8dae59eea944a6faf2edaf5d7a184df2025-08-20T02:15:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0192e8891110.1371/journal.pone.0088911Mitochondrial respiratory function induces endogenous hypoxia.Sara PriorAra KimToshitada YoshiharaSeiji TobitaToshiyuki TakeuchiMasahiro HiguchiHypoxia influences many key biological functions. In cancer, it is generally believed that hypoxic condition is generated deep inside the tumor because of the lack of oxygen supply. However, consumption of oxygen by cancer should be one of the key means of regulating oxygen concentration to induce hypoxia but has not been well studied. Here, we provide direct evidence of the mitochondrial role in the induction of intracellular hypoxia. We used Acetylacetonatobis [2-(2'-benzothienyl) pyridinato-kN, kC3'] iridium (III) (BTP), a novel oxygen sensor, to detect intracellular hypoxia in living cells via microscopy. The well-differentiated cancer cell lines, LNCaP and MCF-7, showed intracellular hypoxia without exogenous hypoxia in an open environment. This may be caused by high oxygen consumption, low oxygen diffusion in water, and low oxygen incorporation to the cells. In contrast, the poorly-differentiated cancer cell lines: PC-3 and MDAMB231 exhibited intracellular normoxia by low oxygen consumption. The specific complex I inhibitor, rotenone, and the reduction of mitochondrial DNA (mtDNA) content reduced intracellular hypoxia, indicating that intracellular oxygen concentration is regulated by the consumption of oxygen by mitochondria. HIF-1α was activated in endogenously hypoxic LNCaP and the activation was dependent on mitochondrial respiratory function. Intracellular hypoxic status is regulated by glucose by parabolic dose response. The low concentration of glucose (0.045 mg/ml) induced strongest intracellular hypoxia possibly because of the Crabtree effect. Addition of FCS to the media induced intracellular hypoxia in LNCaP, and this effect was partially mimicked by an androgen analog, R1881, and inhibited by the anti-androgen, flutamide. These results indicate that mitochondrial respiratory function determines intracellular hypoxic status and may regulate oxygen-dependent biological functions.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0088911&type=printable
spellingShingle Sara Prior
Ara Kim
Toshitada Yoshihara
Seiji Tobita
Toshiyuki Takeuchi
Masahiro Higuchi
Mitochondrial respiratory function induces endogenous hypoxia.
PLoS ONE
title Mitochondrial respiratory function induces endogenous hypoxia.
title_full Mitochondrial respiratory function induces endogenous hypoxia.
title_fullStr Mitochondrial respiratory function induces endogenous hypoxia.
title_full_unstemmed Mitochondrial respiratory function induces endogenous hypoxia.
title_short Mitochondrial respiratory function induces endogenous hypoxia.
title_sort mitochondrial respiratory function induces endogenous hypoxia
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0088911&type=printable
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AT seijitobita mitochondrialrespiratoryfunctioninducesendogenoushypoxia
AT toshiyukitakeuchi mitochondrialrespiratoryfunctioninducesendogenoushypoxia
AT masahirohiguchi mitochondrialrespiratoryfunctioninducesendogenoushypoxia