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Fenretinide induces mitochondrial ROS and inhibits the mitochondrial respiratory chain in neuroblastoma.

Cuperus R, Leen R, Tytgat GA, Caron HN, van Kuilenburg AB - Cell. Mol. Life Sci. (2009)

Bottom Line: ROS induction by fenretinide was of mitochondrial origin, demonstrated by detection of superoxide with MitoSOX, the scavenging effect of the mitochondrial antioxidant MitoQ and reduced ROS production in cells without a functional mitochondrial respiratory chain (Rho zero cells).In digitonin-permeabilized cells, a fenretinide concentration-dependent decrease in ATP synthesis and substrate oxidation was observed, reflecting inhibition of the mitochondrial respiratory chain.Co-incubation of fenretinide with inhibitors of different complexes of the respiratory chain suggested that fenretinide-induced ROS production occurred via complex II.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Genetic Metabolic Diseases, Department of Pediatrics/Emma Children's Hospital, Academic Medical Centre, University of Amsterdam, P.O. Box 22700, 1100 DE, Amsterdam, The Netherlands.

ABSTRACT
Fenretinide induces apoptosis in neuroblastoma by induction of reactive oxygen species (ROS). In this study, we investigated the role of mitochondria in fenretinide-induced cytotoxicity and ROS production in six neuroblastoma cell lines. ROS induction by fenretinide was of mitochondrial origin, demonstrated by detection of superoxide with MitoSOX, the scavenging effect of the mitochondrial antioxidant MitoQ and reduced ROS production in cells without a functional mitochondrial respiratory chain (Rho zero cells). In digitonin-permeabilized cells, a fenretinide concentration-dependent decrease in ATP synthesis and substrate oxidation was observed, reflecting inhibition of the mitochondrial respiratory chain. However, inhibition of the mitochondrial respiratory chain was not required for ROS production. Co-incubation of fenretinide with inhibitors of different complexes of the respiratory chain suggested that fenretinide-induced ROS production occurred via complex II. The cytotoxicity of fenretinide was exerted through the generation of mitochondrial ROS and, at higher concentrations, also through inhibition of the mitochondrial respiratory chain.

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ROS production in Rho zero cells (open square) and control cells (black diamond) measured after 4 h incubation with 0–20 μM 4HPR. ROS was measured using the DFCDA probe. The amount of ROS was expressed relative to that in untreated cells. Each figure represents the mean ± SD of three experiments
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Fig4: ROS production in Rho zero cells (open square) and control cells (black diamond) measured after 4 h incubation with 0–20 μM 4HPR. ROS was measured using the DFCDA probe. The amount of ROS was expressed relative to that in untreated cells. Each figure represents the mean ± SD of three experiments

Mentions: In Rho zero cells, cultured from osteosarcoma cells, the 4HPR-induced ROS production was measured to investigate whether ROS production is due to a specific effect of 4HPR on the respiratory chain. After 4 h incubation with 5 μM 4HPR, hardly any ROS production was observed in Rho zero cells when compared to control osteosarcoma cells (Fig. 4). In addition, 4HPR-induced ROS production, measured using the CM-H2DCFDA probe, was scavenged when the control osteosarcoma cells were pre-incubated for 2 h with 1 μM MitoQ followed by co-incubation with 10 μM 4HPR. Thus, a functional mitochondrial respiratory chain is required for 4HPR-induced ROS production.Fig. 4


Fenretinide induces mitochondrial ROS and inhibits the mitochondrial respiratory chain in neuroblastoma.

Cuperus R, Leen R, Tytgat GA, Caron HN, van Kuilenburg AB - Cell. Mol. Life Sci. (2009)

ROS production in Rho zero cells (open square) and control cells (black diamond) measured after 4 h incubation with 0–20 μM 4HPR. ROS was measured using the DFCDA probe. The amount of ROS was expressed relative to that in untreated cells. Each figure represents the mean ± SD of three experiments
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2824117&req=5

Fig4: ROS production in Rho zero cells (open square) and control cells (black diamond) measured after 4 h incubation with 0–20 μM 4HPR. ROS was measured using the DFCDA probe. The amount of ROS was expressed relative to that in untreated cells. Each figure represents the mean ± SD of three experiments
Mentions: In Rho zero cells, cultured from osteosarcoma cells, the 4HPR-induced ROS production was measured to investigate whether ROS production is due to a specific effect of 4HPR on the respiratory chain. After 4 h incubation with 5 μM 4HPR, hardly any ROS production was observed in Rho zero cells when compared to control osteosarcoma cells (Fig. 4). In addition, 4HPR-induced ROS production, measured using the CM-H2DCFDA probe, was scavenged when the control osteosarcoma cells were pre-incubated for 2 h with 1 μM MitoQ followed by co-incubation with 10 μM 4HPR. Thus, a functional mitochondrial respiratory chain is required for 4HPR-induced ROS production.Fig. 4

Bottom Line: ROS induction by fenretinide was of mitochondrial origin, demonstrated by detection of superoxide with MitoSOX, the scavenging effect of the mitochondrial antioxidant MitoQ and reduced ROS production in cells without a functional mitochondrial respiratory chain (Rho zero cells).In digitonin-permeabilized cells, a fenretinide concentration-dependent decrease in ATP synthesis and substrate oxidation was observed, reflecting inhibition of the mitochondrial respiratory chain.Co-incubation of fenretinide with inhibitors of different complexes of the respiratory chain suggested that fenretinide-induced ROS production occurred via complex II.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Genetic Metabolic Diseases, Department of Pediatrics/Emma Children's Hospital, Academic Medical Centre, University of Amsterdam, P.O. Box 22700, 1100 DE, Amsterdam, The Netherlands.

ABSTRACT
Fenretinide induces apoptosis in neuroblastoma by induction of reactive oxygen species (ROS). In this study, we investigated the role of mitochondria in fenretinide-induced cytotoxicity and ROS production in six neuroblastoma cell lines. ROS induction by fenretinide was of mitochondrial origin, demonstrated by detection of superoxide with MitoSOX, the scavenging effect of the mitochondrial antioxidant MitoQ and reduced ROS production in cells without a functional mitochondrial respiratory chain (Rho zero cells). In digitonin-permeabilized cells, a fenretinide concentration-dependent decrease in ATP synthesis and substrate oxidation was observed, reflecting inhibition of the mitochondrial respiratory chain. However, inhibition of the mitochondrial respiratory chain was not required for ROS production. Co-incubation of fenretinide with inhibitors of different complexes of the respiratory chain suggested that fenretinide-induced ROS production occurred via complex II. The cytotoxicity of fenretinide was exerted through the generation of mitochondrial ROS and, at higher concentrations, also through inhibition of the mitochondrial respiratory chain.

Show MeSH
Related in: MedlinePlus