A comparative study of using comet assay and γH2AX foci formation in the detection of N-methyl-N′-nitro-N-nitrosoguanidine-induced DNA damage

Yu, Yongping; Zhu, Wen; Diao, Huiling; Zhou, Chunxian; Chen, Fanqing F.; Yang, Jing

doi:10.1016/j.tiv.2006.01.004

Cited by 74 publications

(58 citation statements)

References 22 publications

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“…Induction of DNA damage can be determined by the analysis of γH2AX accumulation in the nucleus (Yu et al, 2006). The results from the present study demonstrated that 18α-glycyrrhetinic acid significantly increased the population of HR5-CL11 cells possessing more than four γH2AX foci.…”

Section: Discussionsupporting

confidence: 48%

18α-Glycyrrhetinic acid inhibits the viability of HR5-CL11 cervical carcinoma cells through induction of apoptosis and DNA damage

Zhuang

Hu²,

Hong

et al. 2016

Bangladesh J Pharmacol

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The present study was aimed to investigate the effect of 18α-glycyrrhetinic acid on induction of apoptosis and DNA-damage in HR5-CL11 cervical carcinoma cells. The results revealed 73.6% reduction in HR5-CL11 cell viability on treatment with 5 µM concentration of 18α-glycyrrhetinic acid for 48 hours. The DNA of 18α-glycyrrhetinic acid-treated cells showed a ladder-like pattern. Fragmentation of DNA in the 18α-glycyrrhetinic acid-treated cells was markedly higher compared to the control cells. Examination of the DNA damage in HR5-CL11 cells after treatment with 18α-glycyrrhetinic acid showed breakage in DNA strands and formation of comet-like structures. The frequency of comet formation in 18α-glycyrrhetinic acid treated cells was found to be 7.8 after 48 hours. The population of cells with more than four γH2AX foci was increased to 38.6% on treatment with 5 µM concentration of 18α-glycyrrhetinic acid. Thus, 18α-glycyrrhetinic acid inhibits the viability of HR5-CL11 cervical cancer cells through induction of apoptosis by DNA damage and can be used for the treatment of cervical cancer.Video Clip:<a href="https://www.youtube.com/v/XZQHm07d2sk">MTT assay</a>: 1 min 13 sec

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Section: Discussionsupporting

confidence: 48%

18α-Glycyrrhetinic acid inhibits the viability of HR5-CL11 cervical carcinoma cells through induction of apoptosis and DNA damage

Zhuang

Hu²,

Hong

et al. 2016

Bangladesh J Pharmacol

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“…Among the various types of DNA damage, DSBs are the most severe. ␥H2AX (the phosphorylated form of H2AX) focus formation has been gradually accepted as a sensitive indicator of DSBs (38,39). Hence, A549 cells were infected with 10 CFU/cell of M. pneumoniae and then subjected to immunofluorescence microscopy to evaluate the formation of ␥H2AX foci.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, we further evaluated the generation of reactive oxygen species (ROS) in M. pneumoniae-infected cells. In addition, as ROS are known to induce DNA damage, we also used ␥H2AX focus formation, a new indicator of DNA double-strand breaks (DSBs) (38,39), to determine whether M. pneumoniae can cause DNA damage through the action of ROS. plasma broth base CM403 (Oxoid, Hampshire, United Kingdom), mycoplasma selective supplement G SR59 (Oxoid), 0.5% glucose, and 0.002% phenol red at 37°C in the presence of 5% CO 2 .…”

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Mycoplasma pneumoniae Infection Induces Reactive Oxygen Species and DNA Damage in A549 Human Lung Carcinoma Cells

Sun

Wang

et al. 2008

Infect Immun

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Mycoplasma pneumoniae is a frequent cause of community-acquired bacterial respiratory infections in children and adults. In the present study, using a proteomic approach, we studied the effects of M. pneumoniae infection on the protein expression profile of A549 human lung carcinoma cells. M. pneumoniae infection induced changes in the expression of cellular proteins, in particular a group of proteins involved in the oxidative stress response, such as glucose-6-phosphate 1-dehydrogenase, NADH dehydrogenase (ubiquinone) Fe-S protein 2, and ubiquinol-cytochrome c reductase complex core protein I mitochondrial precursor. The oxidative status of M. pneumoniae-infected cells was evaluated, and the results revealed that M. pneumoniae infection indeed caused generation of reactive oxygen species (ROS). It was further shown that M. pneumoniae infection also induced DNA double-strand breaks, as demonstrated by the formation of ␥H2AX foci. On the other hand, an ROS scavenger, N-acetylcysteine, could inhibit the ROS generation, as well as decrease ␥H2AX focus formation. This is the first report showing that M. pneumoniae infection can directly induce DNA damage, at least partially, through the generation of ROS, and thus this report strengthens the powerful application of proteomics in the study of the pathogenesis of M. pneumoniae.Mycoplasma pneumoniae is one of the smallest self-replicating organisms capable of cell-free existence. As a human pathogen, M. pneumoniae is a major cause of community-acquired respiratory infections in children and adults, which can lead to tracheobronchitis and primary atypical pneumonia (17). Besides causing diseases in the respiratory system, M. pneumoniae has been implicated in several extrapulmonary complications arising from infection; for example, it is a factor in the development of arthritis, cardiovascular diseases, and neurotropic infections (32,34).Considerable progress in our understanding of M. pneumoniae pathogenesis has been made over the years. The activation of the host immune response and direct invasion of cells are believed to contribute to this pathogenesis (32, 37). It has been shown that attachment of M. pneumoniae to the respiratory epithelium is a prerequisite for disease (22). The close interaction between M. pneumoniae and host cells protects the bacterium from removal by the host's mucociliary clearance mechanism and allows it to proliferate and produce metabolites, which in turn can cause cytotoxic effects (32, 34). Simultaneously, M. pneumoniae attachment induces the cells' inflammatory reaction and the host's immune response. For example, upregulation of interlukin-2, -4, -5, -6, -10, -12, and -18 and interferon has been detected in bronchoalveolar lavages, blood, and lungs of M. pneumoniae-infected patients (34). Recently, it was found that the M. pneumoniae protein MPN372, which contains key amino acids similar to the pertussis toxin S1 subunit, might be responsible for airway cellular damage and other sequelae associated with M. pneumoniae infections in human...

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“…Phosphorylation of H2AX is an early event in the ATM-dependent DNA damagesignaling pathway, which has been shown to be active in both myoblasts and myotubes after induction of DSB by ionizing radiation exposure (21). Evidence is accumulating that ␥H2AX foci also identify DNA damage-containing structures other than DSB (22). In the case of myoblasts, ␥H2AX foci were exclusively present in S-phase cells, indicating that they mark the formation of DSB produced by the replication of DNA templates containing SSB.…”

Section: Discussionmentioning

confidence: 99%

Terminally differentiated muscle cells are defective in base excision DNA repair and hypersensitive to oxygen injury

Narciso

Fortini

Pajalunga

et al. 2007

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

107

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The differentiation of skeletal myoblasts is characterized by permanent withdrawal from the cell cycle and fusion into multinucleated myotubes. Muscle cell survival is critically dependent on the ability of cells to respond to oxidative stress. Base excision repair (BER) is the main repair mechanism of oxidative DNA damage. In this study, we compared the levels of endogenous oxidative DNA damage and BER capacity of mouse proliferating myoblasts and their differentiated counterpart, the myotubes. Changes in the expression of oxidative stress marker genes during differentiation, together with an increase in 8-hydroxyguanine DNA levels in terminally differentiated cells, suggested that reactive oxygen species are produced during this process. The repair of 2-deoxyribonolactone, which is exclusively processed by longpatch BER, was impaired in cell extracts from myotubes. The repair of a natural abasic site (a preferred substrate for short-patch BER) also was delayed. The defect in BER of terminally differentiated muscle cells was ascribed to the nearly complete lack of DNA ligase I and to the strong down-regulation of XRCC1 with subsequent destabilization of DNA ligase III␣. The attenuation of BER in myotubes was associated with significant accumulation of DNA damage as detected by increased DNA single-strand breaks and phosphorylated H2AX nuclear foci upon exposure to hydrogen peroxide. We propose that in skeletal muscle exacerbated by free radical injury, the accumulation of DNA repair intermediates, due to attenuated BER, might contribute to myofiber degeneration as seen in sarcopenia and many muscle disorders.oxidative stress ͉ XRCC1 ͉ DNA ligases ͉ DNA single-strand breaks ͉ 8-oxoguanine

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A comparative study of using comet assay and γH2AX foci formation in the detection of N-methyl-N′-nitro-N-nitrosoguanidine-induced DNA damage

Cited by 74 publications

References 22 publications

18α-Glycyrrhetinic acid inhibits the viability of HR5-CL11 cervical carcinoma cells through induction of apoptosis and DNA damage

18α-Glycyrrhetinic acid inhibits the viability of HR5-CL11 cervical carcinoma cells through induction of apoptosis and DNA damage

Mycoplasma pneumoniae Infection Induces Reactive Oxygen Species and DNA Damage in A549 Human Lung Carcinoma Cells

Terminally differentiated muscle cells are defective in base excision DNA repair and hypersensitive to oxygen injury

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