Detection of Radiation-Induced Apoptosis Using the Comet Assay

Wada, Seiichi; Khoa, Tran Van; Kobayashi, Yasuhiko; Funayama, Tomoo; Yamamoto, Kazuo; Natsuhori, Masahiro; Ito, Nobuhiko

doi:10.1292/jvms.65.1161

Cited by 24 publications

(23 citation statements)

References 29 publications

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“…Data from those apoptotic cells would not represent radiation-induced DSBs. Apoptotic cells were differentiated from comet cells in the present study according to the appropriate guidelines [17,18] (Figure 1). We did not include apoptotic cells when we scanned for comet cells under a fluorescence microscope.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of the Comet Assay for Assessing the Dose-Response Relationship of DNA Damage Induced by Ionizing Radiation

Wang

et al. 2013

IJMS

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Dose- and time-response curves were combined to assess the potential of the comet assay in radiation biodosimetry. The neutral comet assay was used to detect DNA double-strand breaks in lymphocytes caused by γ-ray irradiation. A clear dose-response relationship with DNA double-strand breaks using the comet assay was found at different times after irradiation (p < 0.001). A time-response relationship was also found within 72 h after irradiation (p < 0.001). The curves for DNA double-strand breaks and DNA repair in vitro of human lymphocytes presented a nice model, and a smooth, three-dimensional plane model was obtained when the two curves were combined.

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Section: Resultsmentioning

confidence: 99%

Evaluation of the Comet Assay for Assessing the Dose-Response Relationship of DNA Damage Induced by Ionizing Radiation

Wang

et al. 2013

IJMS

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“…Comet assay evaluates apoptotic events of individual cells, whereas DNA ladder and DNA content profiling are used to evaluate cell population. Cells with fan-like tail and small heads in the Comet assay are considered as apoptotic cells (26,27). The comet assay can also evaluate the intensity of DNA damage by computerized analysis of several comet parameters including Lhead, Ltail, head DNA, tail DNA, Lcomet, TM, and OTM (16,26,28).…”

Section: Discussionmentioning

confidence: 99%

Cigarette Smoke Extract Induces DNA Damage but Not Apoptosis in Human Bronchial Epithelial Cells

Liu

Conner

Kobayashi

et al. 2005

Am J Respir Cell Mol Biol

100

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Whether DNA damage caused by cigarette smoke leads to repair or apoptosis has not been fully elucidated. The current study demonstrates that cigarette smoke induces single-strand DNA damage in human bronchial epithelial cells. Cigarette smoke also stimulated caspase 3 precursors as well as intact poly (ADP-ribose) polymerase (PARP) production, but did not activate caspase 3 or cleave PARP, while the alkaloid camptothecin did so. Neither apoptosis nor necrosis was induced by cigarette smoke when the insult was removed within a designated time period. In contrast, DNA damage following cigarette smoke exposure was repaired as evidenced by decreasing terminal dUTP-biotin nick-end labeling positivity. The PARP inhibitor, 3-aminobenzamide blocked this repair. Furthermore, cells subjected to DNA damage were able to survive and proliferate clonogenically when changed to smoke-free conditions. These results suggest that cigarette smoke-induced DNA damage in bronchial epithelial cells is not necessarily lethal, and that PARP functions in the repair process. Our data also suggest that the potency of cigarettes as a carcinogen may result from their ability to induce DNA damage while failing to trigger the apoptotic progression permitting survival of cells harboring potentially oncogenic mutations.

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“…, double-strand breakage, associated with apoptosis and necrosis (Olive et al , 1993; Olive and Banath, 1995; Steinert, 1996; Godard et al , 1999; Tice et al , 2000; Wada et al , 2003). To overcome this limitation, the Comet assay may be used in combination with related methodologies that estimate DNA fragmentation associated with apoptosis and necrosis, such as the DNA diffusion assay (Singh, 2000a,b).…”

Section: Introductionmentioning

confidence: 99%

Estimates of DNA strand breakage in bottlenose dolphin (Tursiops truncatus) leukocytes measured with the Comet and DNA diffusion assays

et al. 2009

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The analysis of DNA damage by mean of Comet or single cell gel electrophoresis (SCGE) assay has been commonly used to assess genotoxic impact in aquatic animals being able to detect exposure to low concentrations of contaminants in a wide range of species. The aims of this work were 1) to evaluate the usefulness of the Comet to detect DNA strand breakage in dolphin leukocytes, 2) to use the DNA diffusion assay to determine the amount of DNA strand breakage associated with apoptosis or necrosis, and 3) to determine the proportion of DNA strand breakage that was unrelated to apoptosis and necrosis. Significant intra-individual variation was observed in all of the estimates of DNA damage. DNA strand breakage was overestimated because a considerable amount (~29%) of the DNA damage was derived from apoptosis and necrosis. The remaining DNA damage in dolphin leukocytes was caused by factors unrelated to apoptosis and necrosis. These results indicate that the DNA diffusion assay is a complementary tool that can be used together with the Comet assay to assess DNA damage in bottlenose dolphins.

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Detection of Radiation-Induced Apoptosis Using the Comet Assay

Cited by 24 publications

References 29 publications

Evaluation of the Comet Assay for Assessing the Dose-Response Relationship of DNA Damage Induced by Ionizing Radiation

Evaluation of the Comet Assay for Assessing the Dose-Response Relationship of DNA Damage Induced by Ionizing Radiation

Cigarette Smoke Extract Induces DNA Damage but Not Apoptosis in Human Bronchial Epithelial Cells

Estimates of DNA strand breakage in bottlenose dolphin (Tursiops truncatus) leukocytes measured with the Comet and DNA diffusion assays

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