Benzene’s metabolites alter c-MYB activity via reactive oxygen species in HD3 cells☆

Wan, Joanne; Winn, Louise M.

doi:10.1016/j.taap.2007.04.016

Cited by 25 publications

(21 citation statements)

References 58 publications

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“…The DCF assay is a popular method for assessing toxicant-stimulated generation of reactive oxidant species in cells (Bussche & Soares, 2011; Filipič & Hei, 2004; Gong & Han, 2006; Hatcher, et al, 2008; Hu, et al, 2011; Michalowicz, 2010; Naqvi, et al, 2010; Shanker & Aschner, 2003; Shao, et al, 2008; Wan & Winn, 2007; Zhang, et al, 2011; Zhu, et al, 2009). Although numerous caveats have been reported with the DCF assay, including effects of medium composition, serum, heme, heme proteins, metalloporphyrins and bovine serum albumin (Chen, et al, 2010), interactions with chemical toxicants have received nominal attention.…”

Section: Discussionmentioning

confidence: 99%

Troubleshooting the dichlorofluorescein assay to avoid artifacts in measurement of toxicant-stimulated cellular production of reactive oxidant species

Tetz

Kamau

Cheng

et al. 2013

Journal of Pharmacological and Toxicological Methods

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Introduction The dichlorofluorescein (DCF) assay is a popular method for measuring cellular reactive oxidant species (ROS). Although caveats have been reported with the DCF assay and other compounds, the potential for artifactual results due to cell-free interactions between the DCF compound and toxicants has hardly been explored. We evaluated the utility of the DCF assay for measuring ROS generation by the toxicants mono-(2-ethylhexyl) phthalate (MEHP), and tetrabromobisphenol A (TBBPA). Methods DCF fluorescence was measured spectrofluorometrically after a 1-h incubation of toxicants with 6-carboxy-2′,7′-dichlorodihydrofluorescein diacetate (carboxy-H2DCFDA). MEHP was incubated with carboxy-H2DCFDA in cell-free solutions of Hank’s buffered salt solution (HBSS), or in Royal Park Memorial Institute (RPMI) medium with or without fetal bovine serum. TBBPA was incubated with carboxy-H2DCFDA in cell-free HBSS and with human trophoblast cells (HTR8/SVneo cells). Results MEHP did not increase fluorescence in solutions of carboxy-H2DCFDA in HBSS or RPMI medium without serum. However, MEHP (90 and 180 μM) increased DCF fluorescence in cell-free RPMI medium containing serum. Furthermore, serum-free and cell-free HBSS solutions containing 25 μM TBBPA exhibited concentration-dependent increased fluorescence with 5–100 μM carboxy-H2DCFDA (p<0.05), but not 1 μM carboxy-H2DCFDA. In addition, we observed increased fluorescence in HTR8/SVneo cell cultures exposed to TBBPA (0.5–25 μM) (p<0.05), as we had observed in cell-free buffer. Discussion MEHP demonstrated an interaction with serum in cell-free generation of DCF fluorescence, whereas TBBPA facilitated conversion of carboxy-H2DCFDA to the fluorescent DCF moiety in the absence of serum. Because TBBPA increased fluorescence in the absence of cells, the increased DCF fluorescence observed with TBBPA in the presence of cells cannot be attributed to cellular ROS and may, instead, be the result of chemical activation of carboxy-H2DCFDA to the fluorescent DCF moiety. These data illustrate the importance of including cell-free controls when using the DCF assay to study toxicant-stimulated cellular production of ROS.

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Section: Discussionmentioning

confidence: 99%

Troubleshooting the dichlorofluorescein assay to avoid artifacts in measurement of toxicant-stimulated cellular production of reactive oxidant species

Tetz

Kamau

Cheng

et al. 2013

Journal of Pharmacological and Toxicological Methods

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“…In previous study, 1,4-BQ induced ROS in chicken erythroblast HD3 cells. It is suggested that benzene may exert its toxicity through ROS-mediated c-myb signaling pathway in vitro [7]. 1,4-BQ also forms adducts of DNA or protein in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

Proteome Changes of Human Bone Marrow Mesenchymal Stem Cells Induced by 1,4-Benzoquinone

Zhang

et al. 2016

BioMed Research International

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Benzene is metabolized to hydroquinone in liver and subsequently transported to bone marrow for further oxidization to 1,4-benzoquinone (1,4-BQ), which may be related to the leukemia and other blood disorders. In the present study, we investigated the proteome profiles of human primary bone marrow mesenchymal stem cells (hBM-MSCs) treated by 1,4-BQ. We identified 32 proteins that were differentially expressed. Two of them, HSP27 and Vimentin, were verified at both mRNA and protein levels and their cellular localization was examined by immunofluorescence. We also found increased mRNA level of RAP1GDS1, a critical factor of metabolism that has been identified as a fusion partner in various hematopoietic malignancies. Therefore, these differentially expressed proteins can play important roles in benzene-mediated hematoxicity.

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“…Benzene can be bioactivated to toxic metabolites such as 1,4 benzoquinone, which can alter signalling pathways and affect chromosomal integrity. Benzoquinone has been shown to increase the activity of c-Myb, which is an important transcription factor involved in haematopoiesis, cell proliferation and cell differentiation 1. A recent study has also shown that exposure to low-dose benzene (urban traffic police) could alter DNA methylation and induce epigenetic changes in gene expression 2…”

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confidence: 99%

Acute childhood leukaemia and residence next to petrol stations and automotive repair garages: the ESCALE study (SFCE)

et al. 2009

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Benzene’s metabolites alter c-MYB activity via reactive oxygen species in HD3 cells☆

Cited by 25 publications

References 58 publications

Troubleshooting the dichlorofluorescein assay to avoid artifacts in measurement of toxicant-stimulated cellular production of reactive oxidant species

Troubleshooting the dichlorofluorescein assay to avoid artifacts in measurement of toxicant-stimulated cellular production of reactive oxidant species

Proteome Changes of Human Bone Marrow Mesenchymal Stem Cells Induced by 1,4-Benzoquinone

Acute childhood leukaemia and residence next to petrol stations and automotive repair garages: the ESCALE study (SFCE)

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