Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay

Atha, Donald H.; Coşkun, Erdem; Erdem, Onur; Tona, Alessandro; Reipa, Vytas; Nelson, Bryant C.

doi:10.1155/2020/8810105

Cited by 14 publications

(20 citation statements)

References 32 publications

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“…The use of suspended cells could alleviate the problem of insufficient cells for comet analysis at high treatment levels. FreeStyle cell preparations would also be well suited for additional characterization methods that require isolation of DNA, such as mass spectrometry [ 43 ]. For the preparation of a reference material, the treated cells would also need to be quickly frozen and stored at −150°C to prevent DNA repair before they are used for subsequent certification by the comet and viability assay and any additional measurements [ 43 ].…”

Section: Resultsmentioning

confidence: 99%

“…FreeStyle cell preparations would also be well suited for additional characterization methods that require isolation of DNA, such as mass spectrometry [ 43 ]. For the preparation of a reference material, the treated cells would also need to be quickly frozen and stored at −150°C to prevent DNA repair before they are used for subsequent certification by the comet and viability assay and any additional measurements [ 43 ]. Certification by multiple methods of comet analysis such as %tail DNA and OTM would be useful.…”

Section: Resultsmentioning

confidence: 99%

“…Within about 5 min. following UVA exposure, the control and treated cells were harvested by trypsin EDTA (2.5 mg/mL) and measured by alkaline comet assay as described previously [41][42][43] and again as follows. Low-melting point agarose, 300 μL (Trevigen Inc., MD, USA, cat.…”

Section: Methodsmentioning

confidence: 99%

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Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability

Atha

Tona

Reipa

2022

Journal of Nucleic Acids

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Application of DNA damage diagnostic tests is rapidly growing, in particular for ovarian, prostate, and skin cancers; environmental monitoring; chronic and degenerative diseases; and male infertility. Such tests suffer from significant variability among different laboratories due the lack of standardization, experimental validation, and differences in data interpretation. Reference methods and materials for quantitative measurement of UVA-induced DNA damage in mammalian cells are frequently needed. In this study, we examined the use of the single-cell gel electrophoresis (comet) assay to assess the UVA-induced DNA damage in surface-attached Chinese hamster ovary (CHO) cells treated with a photosensitizer as a candidate cellular oxidative damage reference material. We found that the comet images became diffused and the viability of the cells decreased substantially (>20%) as the UVA dose and benzo [a] pyrene (BaP) concentration exceeded 6.3 J/cm 2 and 10 −6 mol/L BaP. Maintaining the conditions of exposure within this range can improve DNA damage measurement fidelity, particularly if used as a quantitative reference method and to produce materials considered as an in vitro standard for the comet assay.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability

Atha

Tona

Reipa

2022

Journal of Nucleic Acids

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“…The aim of this study was to examine the effect of these compounds on normal cells as they have the potential for systemic treatment of various, non-cancerous diseases. Chinese hamster ovary (CHO) cells were chosen, as they represent a widely used model system for the comet assay [21][22][23], which are easy to handle and highly tolerant to variations in pH, oxygen levels and temperature. Since there are no data about the potential cytotoxicity and genotoxicity of these novel compounds, here we investigated their effects on cell viability and DNA stability.…”

Section: Introductionmentioning

confidence: 99%

Synthetic Diphenylacetylene-Based Retinoids Induce DNA Damage in Chinese Hamster Ovary Cells without Altering Viability

et al. 2022

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All-trans-retinoic acid (ATRA), the active metabolite of vitamin A, plays a pivotal role in cell differentiation, proliferation and embryonic development. It is an effective therapy for dermatological disorders and malignancies. ATRA is prone to isomerization and oxidation, which can affect its activity and selectivity. Novel diphenylacetylene-based ATRA analogues with increased stability can help to overcome these problems and may offer significant potential as therapeutics for a variety of cancers and neurodegenerative diseases, including amyotrophic lateral sclerosis. Here, we investigated the effects of these retinoids on cell viability and genotoxicity in the widely used model system of the rapidly proliferating Chinese hamster ovary cell line. DC360 is a fluorescent ATRA analogue and DC324 is a non-active derivative of DC360. EC23, DC525, DC540, DC645, and DC712 are promising analogues with increased bioactivity. The cytotoxic activity of the compounds was evaluated by ATP assay and DNA damage was tested by comet assay. No cytotoxicity was observed in the 10−6–10−5 M concentration range. All compounds induced DNA migration similar to ATRA, but DC324, DC360 and EC23 did so to a greater extent, particularly at higher concentrations. We believe that retinoid receptor-independent genotoxicity is a general characteristic of these compounds; however, further studies are needed to identify the molecular mechanisms and understand their complex biological functions.

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“…Several methods have been reported in the literature for the estimation of ETO, MOX and NAL in their pharmaceutical dosage form and/or biological fluids; for ETO: spectrophotometric [8], spectrofluorimetric (SF) [9], electrochemical [10,11], high-performance thin-layer chromatography (HPTLC) [12], high-performance liquid chromatography (HPLC) [13][14][15][16][17], gas chromatography (GC) [18] and capillary electrophoresis (CP) [19]; for MOX: spectrophotometric [20,21], SF [22], electrochemical [23], HPTLC [24], HPLC [25][26][27] and CP [28] and for NAL spectrophotometric [29][30][31], SF [31,32], flow injection analysis [33,34], HPLC [35][36][37] and GC [38]. These presented methods did not guide the monitoring of these three drugs in the biological matrices.…”

Section: Introductionmentioning

confidence: 99%

Eco-friendly fluorimetric approaches for the simultaneous estimation of the co-administered ternary mixture: etoposide, moxifloxacin and nalbuphine

2021

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Antineoplastic drugs, etoposide (ETO), are widely used in leukaemia. A patient with leukaemia has a relative infection with pneumonia treated by fluoroquinolones as moxifloxacin HCL (MOX). Because opioid analgesic as nalbuphine HCL (NAL) does not have a ceiling dose, it is used to manage the distasteful sensory in leukaemia. Consequently, green methods for synchronous spectrofluorimetric quantification of a ternary mixture of ETO, MOX and NAL were developed. The first approach relies simply on the estimation of MOX at 371 nm by conventional synchronous fluorimetric technique (Δ λ of 60 nm). The second approach depends on applying the first derivative synchronous fluorimetric technique (Δ λ of 60 nm) for simultaneous estimation of ETO and NAL at 257 and 273 nm, respectively. A good linear correlation was obtained in the ranges of 0.04–0.40, 0.10–1.00 and 0.50–5.00 µg ml −1 for MOX, ETO and NAL, respectively. Moreover, the proposed approaches were successfully applied for the estimation of the studied drugs in the pharmaceutical dosage forms. Additionally, the synchronous assessment of ETO, MOX and NAL in the spiked human urine was successfully attained by the facile protein precipitation technique. The mean % recoveries in spiked human urine were 99.49, 98.07 and 98.48 for MOX, ETO and NAL, respectively.

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Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay

Cited by 14 publications

References 32 publications

Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability

Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability

Synthetic Diphenylacetylene-Based Retinoids Induce DNA Damage in Chinese Hamster Ovary Cells without Altering Viability

Eco-friendly fluorimetric approaches for the simultaneous estimation of the co-administered ternary mixture: etoposide, moxifloxacin and nalbuphine

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