Role of H2O2 in <i>RET/PTC1</i> Chromosomal Rearrangement Produced by Ionizing Radiation in Human Thyroid Cells

Ameziane-El-Hassani, Rabii; Boufraqech, Myriem; Lagente-Chevallier, Odile; Weyemi, Urbain; Talbot, Monique; Métivier, Didier; Courtin, Françoise; Bidart, Jean Michel; Mzibri, Mohammed El; Schlumberger, Martin; Dupuy, Corinne

doi:10.1158/0008-5472.can-09-4336

Cited by 78 publications

(44 citation statements)

References 41 publications

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“…Exogenous H 2 O 2 induces DNA damage in thyroid cells (6,20). To establish a link between H 2 O 2 -generating NADPH oxidase DUOX1 activation and DNA damage observed several days after irradiation, we performed RNA interference experiments ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Hypoxia and antioxidant therapy reduce the X-ray-induced delayed effects, suggesting that radio-induced oxidative stress plays a significant role in determining the susceptibility of irradiated cells to genetic instability (3)(4)(5). We recently showed that H 2 O 2 is able to cause RET/PTC1 rearrangement in thyroid cells, indicating that oxidative stress could be responsible for the RET/PTC rearrangement frequently found in radiation-induced thyroid tumors (6).…”

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confidence: 99%

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NADPH oxidase DUOX1 promotes long-term persistence of oxidative stress after an exposure to irradiation

Ameziane-El-Hassani

Talbot

Santos

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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109

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Ionizing radiation (IR) causes not only acute tissue damage, but also late effects in several cell generations after the initial exposure. The thyroid gland is one of the most sensitive organs to the carcinogenic effects of IR, and we have recently highlighted that an oxidative stress is responsible for the chromosomal rearrangements found in radio-induced papillary thyroid carcinoma. Using both a human thyroid cell line and primary thyrocytes, we investigated the mechanism by which IR induces the generation of reactive oxygen species (ROS) several days after irradiation. We focused on NADPH oxidases, which are specialized ROS-generating enzymes known as NOX/DUOX. Our results show that IR induces delayed NADPH oxidase DUOX1-dependent H 2 O 2 production in a dose-dependent manner, which is sustained for several days. We report that p38 MAPK, activated after IR, increased DUOX1 via IL-13 expression, leading to persistent DNA damage and growth arrest. Pretreatment of cells with catalase, a scavenger of H 2 O 2 , or DUOX1 down-regulation by siRNA abrogated IR-induced DNA damage. Analysis of human thyroid tissues showed that DUOX1 is elevated not only in human radio-induced thyroid tumors, but also in sporadic thyroid tumors. Taken together, our data reveal a key role of DUOX1-dependent H 2 O 2 production in long-term persistent radio-induced DNA damage. Our data also show that DUOX1-dependent H 2 O 2 production, which induces DNA double-strand breaks, can cause genomic instability and promote the generation of neoplastic cells through its mutagenic effect.ionizing radiation | oxidative stress | NADPH oxidase | thyroid |

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

NADPH oxidase DUOX1 promotes long-term persistence of oxidative stress after an exposure to irradiation

Ameziane-El-Hassani

Talbot

Santos

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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109

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“…Because of its numerous advantageous properties, HyPer holds promise for understanding the biological roles of H 2 O 2 , and its signal has been characterized to an extent and used in many different contexts [15][16][17][18][19][20][21].…”

Section: Introductionmentioning

confidence: 99%

In-depth characterization of the fluorescent signal of HyPer, a probe for hydrogen peroxide, in bacteria exposed to external oxidative stress

Lim

Barker

Huang

et al. 2014

Journal of Microbiological Methods

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Highlights• We characterized the signal of HyPer, a fluorescent probe for peroxide, in E. coli.• Each strain requires its own specific signal characterization.• HyPer's signal is reversible rather than real-time.• Expression of HyPer reduced the rate of peroxide scavenging by the expression host.• Careful, controlled use of HyPer facilitates quantitative comparisons across studies. AbstractGenetically encoded, fluorescent biosensors have been developed to probe the activities of various signaling molecules inside cells ranging from changes in intracellular ion concentrations to dynamics of lipid second messengers. HyPer is a member of this class of biosensors and is the first to dynamically respond to hydrogen peroxide (H 2 O 2 ), a reactive oxygen species that functions as a signaling molecule. However, detailed characterization of HyPer's signal is not currently available within the context of bacteria exposed to external oxidative stress, which occurs in the immunological response of higher organisms against invasive pathogenic bacteria. Here, we performed this characterization, specifically in Escherichia coli exposed to external H 2 O 2 . We found that the temporal behavior of the signal does not correspond exactly to peroxide concentration in the system as a function of time and expression of the sensor decreases the peroxide scavenging activity of the cell. We also determined the effects of cell number, both before and after normalization of externally added H 2 O 2 to the number of cells. Finally, we report quantitative characteristics of HyPer's signal in this context, including the dynamic range of the signal, the signal-to-noise ratio, and the half saturation constant. These parameters show statistically meaningful differences in signal between two commonly used strains of E. coli, demonstrating how signal can vary with strain. Taken together, our results establish a systematic, quantitative framework for researchers seeking to better understand the role of H 2 O 2 in the immunological response against bacteria, and for understanding potential differences in the details of HyPer's quantitative performance across studies.

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“…This finding could be explained by the significant decrease in RET/PTC over the years and the equivalent increasing rate of BRAFV600E and RAS mutations in PTC, possibly attributed to the decreased exposure to ionizing radiation in the last decades or to new pollutants (75,76,77,78,79).…”

Section: Discussionmentioning

confidence: 99%

Break–apart interphase fluorescence in situ hybridization assay in papillary thyroid carcinoma: on the road to optimizing the cut-off level for RET/PTC rearrangements

Colato

Vicentini

Cantara

et al. 2015

European Journal of Endocrinology

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Objective: Chromosomal rearrangements of the RET proto-oncogene is one of the most common molecular events in papillary thyroid carcinoma (PTC). However, their pathogenic role and clinical significance are still debated. This study aimed to investigate the prevalence of RET/PTC rearrangement in a cohort of BRAF WT PTCs by fluorescence in situ hybridization (FISH) and to search a reliable cut-off level in order to distinguish clonal or non-clonal RET changes. Design: Forty BRAF WT PTCs were analyzed by FISH for RET rearrangements. As controls, six BRAFV600E mutated PTCs, 13 follicular adenomas (FA), and ten normal thyroid parenchyma were also analyzed. Methods: We performed FISH analysis on formalin-fixed, paraffin-embedded tissue using a commercially available RET break-apart probe. A cut-off level equivalent to 10.2% of aberrant cells was accepted as significant. To validate FISH results, we analyzed the study cohort by qRT-PCR. Results: Split RET signals above the cut-off level were observed in 25% (10/40) of PTCs, harboring a percentage of positive cells ranging from 12 to 50%, and in one spontaneous FA (1/13, 7.7%). Overall, the data obtained by FISH matched well with qRT-PCR results. Challenging findings were observed in five cases showing a frequency of rearrangement very close to the cut-off. Conclusions: FISH approach represents a powerful tool to estimate the ratio between broken and non-broken RET tumor cells. Establishing a precise FISH cut-off may be useful in the interpretation of the presence of RET rearrangement, primarily when this strategy is used for cytological evaluation or for targeted therapy.

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Role of H2O2 in RET/PTC1 Chromosomal Rearrangement Produced by Ionizing Radiation in Human Thyroid Cells

Cited by 78 publications

References 41 publications

NADPH oxidase DUOX1 promotes long-term persistence of oxidative stress after an exposure to irradiation

NADPH oxidase DUOX1 promotes long-term persistence of oxidative stress after an exposure to irradiation

In-depth characterization of the fluorescent signal of HyPer, a probe for hydrogen peroxide, in bacteria exposed to external oxidative stress

Break–apart interphase fluorescence in situ hybridization assay in papillary thyroid carcinoma: on the road to optimizing the cut-off level for RET/PTC rearrangements

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