Whole cell-ELISA to measure the γH2AX response of six aneugens and eight DNA-damaging chemicals

Matsuzaki, Kaori; Harada, Asako; Takeiri, Akira; Tanaka, Kenji; Mishima, Masayuki

doi:10.1016/j.mrgentox.2010.05.009

Cited by 42 publications

(25 citation statements)

References 43 publications

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“…The results showed that 5-FU, CDDP and ET, which belong to the classical anti-cancer drugs that directly interact with DNA function, induced formation of γH2AX foci. Similar observations were reported also by other researchers, who demonstrated that 5-FU (Ikeda et al 2010;Matsuzaki et al 2010), CDDP (Clingen et al 2008;Huang et al 2004;Ikeda et al 2010;Matsuzaki et al 2010;Olive and Banath 2009) and ET (Smart et al 2008;Matsuzaki et al 2010;Muslimovic et al 2009) can induce phosphorylation of histone H2AX in different cell lines. All three drugs also induce chromosomal aberrations in vitro (Gajski et al 2015).…”

Section: Discussionsupporting

confidence: 89%

Influence of selected anti-cancer drugs on the induction of DNA double-strand breaks and changes in gene expression in human hepatoma HepG2 cells

Novak

Žegura

Baebler

et al. 2015

Environ Sci Pollut Res

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In chemotherapy, various anti-cancer drugs with different mechanisms of action are used and may represent different risk of undesirable delayed side effects in treated patients as well as in occupationally exposed populations. The aim of the present study was to evaluate genotoxic potential of four widely used anti-cancer drugs with different mechanisms of action: 5-fluorouracil (5-FU), cisplatin (CDDP) and etoposide (ET) that cause cell death by targeting DNA function and imatinib mesylate (IM) that inhibits targeted protein kinases in cancer cells in an experimental model with human hepatoma HepG2 cells. After 24 h of exposure all four anti-cancer drugs at non-cytotoxic concentrations induced significant increase in formation of DNA double strand breaks (DSBs), with IM being the least effective. The analysis of the changes in the expression of genes involved in the response to DNA damage (CDKN1A, GADD45A, MDM2), apoptosis (BAX, BCL2) and oncogenesis (MYC, JUN) showed that 5-FU, CDDP and ET upregulated the genes involved in DNA damage response, while the anti-apoptotic gene BCL2 and oncogene MYC were downregulated. On the contrary, IM did not change the mRNA level of the studied genes, showing different mechanism of action that probably does not involve direct interaction with DNA processing. Genotoxic effects of the tested anti-cancer drugs were observed at their therapeutic concentrations that may consequently lead to increased risk for development of delayed adverse effects in patients. In addition, considering the genotoxic mechanism of action of 5-FU, CDDP and ET an increased risk can also not be excluded in occupationally exposed populations. The results also indicate that exposure to 5-FU, CDDP and ET represent a higher risk for delayed effects such as cancer, reproductive effects and heritable disease than exposure to IM.

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Section: Discussionsupporting

confidence: 89%

Influence of selected anti-cancer drugs on the induction of DNA double-strand breaks and changes in gene expression in human hepatoma HepG2 cells

Novak

Žegura

Baebler

et al. 2015

Environ Sci Pollut Res

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“…It was also reported that micronucleus formation is correlated with H2AX phosphorylation [7] and that ␥H2AX is a reliable biomarker of pre-cancerous cells in vivo [8,9]. These data support the assumption that H2AX phosphorylation could be an appropriate biomarker of genotoxicity, as suggested in recent in vitro and in vivo studies [10][11][12][13][14][15][16][17][18][19].…”

Section: Introductionsupporting

confidence: 79%

Evidence of the in vitro genotoxicity of methyl-pyrazole pesticides in human cells

Graillot

Tomasetig

Cravedi

et al. 2012

Mutation Research/Genetic Toxicology and Environmental Mutagene

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Consumers are exposed daily to several pesticide residues in food, which can be of potential concern for human health. Based on a previous study dealing with exposure of the French population to pesticide residues via the food, we selected 14 pesticides frequently found in foodstuffs, on the basis of their persistence in the environment or their bioaccumulation in the food chain. In a first step, the objective of this study was to investigate if the 14 selected pesticides were potentially cytotoxic and genotoxic. For this purpose, we used a new and sensitive genotoxicity assay (the γH2AX test, involving phosphorylation of histone H2AX) with four human cell lines (ACHN, SH-SY5Y, LS-174T and HepG2), each originating from a potential target tissue of food contaminants (kidney, nervous system, colon, and liver, respectively). Tebufenpyrad was the only compound identified as genotoxic and the effect was only observed in the SH-SY5Y neuroblastoma cell-line. A time-course study showed that DNA damage appeared early after treatment (1h), suggesting that oxidative stress could be responsible for the induction of γH2AX. In a second step, three other pesticides were studied, i.e. bixafen, fenpyroximate and tolfenpyrad, which - like tebufenpad - also had a methyl-pyrazole structure. All these compounds demonstrated genotoxic activity in SH-SY5Y cells at low concentration (nanomolar range). Complementary experiments demonstrated that the same compounds show genotoxicity in a human T-cell leukemia cell line (Jurkat). Moreover, we observed an increased production of reactive oxygen species in Jurkat cells in the presence of the four methyl-pyrazoles. These results demonstrate that tebufenpyrad, bixafen, fenpyroximat and tolfenpyrad induce DNA damage in human cell lines, very likely by a mode of action that involves oxidative stress. Nonetheless, additional in vivo data are required before a definitive conclusion can be drawn regarding hazard prediction to humans.

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“…Representative images of treated A549 cells are shown in Figure 7. VP-16 is known to cause DNA DSBs and γH2AX has been shown to be a sensitive marker of DNA DSBs induced by a variety of DNA damaging agents including VP-16 [42,43,44,45]. In the present study, γH2AX foci (red channel) were indeed observed when cells were treated with VP-16 (row #2).…”

Section: Resultssupporting

confidence: 62%

Combination of Near Infrared Light-Activated Photodynamic Therapy Mediated by Indocyanine Green with Etoposide to Treat Non-Small-Cell Lung Cancer

Luo

Zhang

Lü

2017

Cancers

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Indocyanine green (ICG) has been reported as a potential near-infrared (NIR) photosensitizer for photodynamic therapy (PDT) of cancer. However the application of ICG-mediated PDT is both intrinsically and physiologically limited. Here we report a combination of ICG-PDT with a chemotherapy drug etoposide (VP-16), aiming to enhance the anticancer efficacy, to circumvent limitations of PDT using ICG, and to reduce side effects of VP-16. We found in controlled in vitro cell-based assays that this combination is effective in killing non-small-cell lung cancer cells (NSCLC, A549 cell line). We also found that the combination of ICG-PDT and VP-16 exhibits strong synergy in killing non-small-cell lung cancer cells partially through inducing more DNA double-strand breaks (DSBs), while it has a much weaker synergy in killing human normal cells (GM05757). Furthermore, by studying the treatment sequence dependence and the cytotoxicity of laser-irradiated mixtures of ICG and VP-16, we found that the observed synergy involves direct/indirect reactions between ICG and VP-16. We further propose that there exists an electron transfer reaction between ICG and VP-16 under irradiation. This study therefore shows the anticancer efficacy of ICG-PDT combined with VP-16. These findings suggest that ICG-mediated PDT may be applied in combination with the chemotherapy drug VP-16 to treat some cancers, especially the non-small-cell lung cancer.

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Whole cell-ELISA to measure the γH2AX response of six aneugens and eight DNA-damaging chemicals

Cited by 42 publications

References 43 publications

Influence of selected anti-cancer drugs on the induction of DNA double-strand breaks and changes in gene expression in human hepatoma HepG2 cells

Influence of selected anti-cancer drugs on the induction of DNA double-strand breaks and changes in gene expression in human hepatoma HepG2 cells

Evidence of the in vitro genotoxicity of methyl-pyrazole pesticides in human cells

Combination of Near Infrared Light-Activated Photodynamic Therapy Mediated by Indocyanine Green with Etoposide to Treat Non-Small-Cell Lung Cancer

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