Optimization of a pre-metabolization procedure using rat liver S9 and cell-extracted S9 in the Ames fluctuation test

Shao, Ying; Schiwy, Andreas; Glauch, Lisa; Henneberger, Luise; König, Maria; Mühlenbrink, Marie; Xiao, Huang; Thalmann, Beat; Schlichting, Rita; Hollert, Henner; Escher, Beate I.

doi:10.1016/j.scitotenv.2020.141468

Cited by 14 publications

(3 citation statements)

References 45 publications

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“…These results demonstrate that MSSV impeded the metastatic ability of HCT116 cells via MMP-9 inhibition mediated by reduced the binding activity of transcription factors. The toxicity of the parent compounds can be determined only if metabolic enzymes to convert the parent compound to metabolites are present in an in vitro system (Chitrangi et al, 2017;Shao et al, 2020). The liver is considered a major site of metabolism because metabolism centered on the liver has been studied, including primary cultures of hepatocytes, subcellular fractions, precision-cut liver tissue slices and drug-metabolizing enzymes (Rodrigues, 1994).…”

Section: Discussionmentioning

confidence: 99%

N-methylsansalvamide elicits antitumor effects in colon cancer cells in vitro and in vivo by regulating proliferation, apoptosis, and metastatic capacity

2023

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N-methylsansalvamide (MSSV), a cyclic pentadepsipeptide, was obtained from a strain of Fusarium solani f. radicicola. The current study investigated the anti-colorectal cancer effect of MSSV. MSSV exhibited the inhibition of the proliferation in HCT116 cells via induction of G0/G1 cell cycle arrest by downregulating CDK 2, CDK6, cyclin D, and cyclin E, and upregulating p21WAF1 and p27KIP1. Decreased phosphorylation of AKT was observed in MSSV-treated cells. Moreover, MSSV treatment induced caspase-mediated apoptosis through elevating the level of cleaved caspase 3, cleaved PARP, cleaved caspase 9, and pro-apoptotic Bax. MSSV revealed the declined MMP-9 level mediated by reduction in the binding activity of AP-1, Sp-1, and NF-κB motifs, which led to the migration and invasion of HCT116 cells. In vitro metabolism with rat liver S9 fractions was performed to examine the effect of MSSV metabolites. The metabolic process enhanced the inhibitory effect of MSSV on the HCT116 cell proliferation via decline of cyclin D1 expression and AKT phosphorylation. Finally, oral administration of MSSV inhibited the tumor growth of HCT116 xenograft mice. These results suggest that MSSV is a potential anti-tumor agent in colorectal cancer treatment.

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Section: Discussionmentioning

confidence: 99%

N-methylsansalvamide elicits antitumor effects in colon cancer cells in vitro and in vivo by regulating proliferation, apoptosis, and metastatic capacity

2023

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“…However, the present dataset indicates that Aroclor 1254-induced S9, which will not be available anymore in the near future, can likely be adequately replaced by PB/β-NF induced S9, without any anticipated negative impact on the LEC values. In the long term an animal-free S9 source may become preferable [ 46 ]. While initial results looks highly promising [ 44 ], more data needs to be provided, before such material can be considered a valid alternative.…”

Section: Discussionmentioning

confidence: 99%

Direct Comparison of the Lowest Effect Concentrations of Mutagenic Reference Substances in Two Ames Test Formats

Rainer

Pinter

Prielinger

et al. 2021

Toxics

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The Ames assay is the standard assay for identifying DNA-reactive genotoxic substances. Multiple formats are available and the correct choice of an assay protocol is essential for achieving optimal performance, including fit for purpose detection limits and required screening capacity. In the present study, a comparison of those parameters between two commonly used formats, the standard pre-incubation Ames test and the liquid-based Ames MPF™, was performed. For that purpose, twenty-one substances with various modes of action were chosen and tested for their lowest effect concentrations (LEC) with both tests. In addition, two sources of rat liver homogenate S9 fraction, Aroclor 1254-induced and phenobarbital/β-naphthoflavone induced, were compared in the Ames MPF™. Overall, the standard pre-incubation Ames and the Ames MPF™ assay showed high concordance (>90%) for mutagenic vs. non-mutagenic compound classification. The LEC values of the Ames MPF™ format were lower for 17 of the 21 of the selected test substances. The S9 source had no impact on the test results. This leads to the conclusion that the liquid-based Ames MPF™ assay format provides screening advantages when low concentrations are relevant, such as in the testing of complex mixtures.

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“…In vitro metabolism bioassays have been proposed as a potential means to expand the capabilities of wastewater-based epidemiology (Lai et al, 2015;Lopardo et al, 2017;Reid et al, 2014). For example, external metabolization systems like the liver S9 fraction in vitro bioassay have been developed (Shao et al, 2020) and refined for high-throughput application (Escher et al, 2020;Huber et al, 2021;Villeneuve et al, 2019). Metabolism is also an important determinant of toxicity since chemicals can be activated or deactivated (detoxified) by, for example, Phase 1 cytochrome P450 monooxygenase (CYP) enzymes (Yu, 2020) and https://doi.org/10.26434/chemrxiv-2023-mvc6g ORCID: https://orcid.org/0000-0003-2724-9183 Content not peer-reviewed by ChemRxiv.…”

Section: Introductionmentioning

confidence: 99%

Improving Wastewater-Based Epidemiology for New Psychoactive Substance Surveillance by Combining a High-Throughput In Vitro Metabolism Assay and LC−HRMS Metabolite Identification

Bade,

Huchthausen,

Huber

et al. 2023

Preprint

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One of the primary criteria for a suitable drug biomarker for wastewater-based epidemiology (WBE) is having a unique source representing human metabolism. For WBE studies, this means it is important to identify and monitor metabolites rather than parent drugs, to capture consumption of drugs and not fractions that could be directly disposed. In this study, a high-throughput workflow based on a human liver S9 fraction in vitro metabolism assay was developed to identify human transformation products of new chemicals, using α-pyrrolidino-2-phenylacetophenone (α-D2PV) as a case study. Analysis by liquid chromatography coupled to high resolution mass spectrometry identified four metabolites. Subsequently, a targeted liquid chromatography – tandem mass spectrometry method was developed for their analysis in wastewater samples collected from a music festival in Australia. The successful application of this workflow opens the door for future work to better understand the metabolism of chemicals and their detection and application for wastewater-based epidemiology.

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Optimization of a pre-metabolization procedure using rat liver S9 and cell-extracted S9 in the Ames fluctuation test

Cited by 14 publications

References 45 publications

N-methylsansalvamide elicits antitumor effects in colon cancer cells in vitro and in vivo by regulating proliferation, apoptosis, and metastatic capacity

N-methylsansalvamide elicits antitumor effects in colon cancer cells in vitro and in vivo by regulating proliferation, apoptosis, and metastatic capacity

Direct Comparison of the Lowest Effect Concentrations of Mutagenic Reference Substances in Two Ames Test Formats

Improving Wastewater-Based Epidemiology for New Psychoactive Substance Surveillance by Combining a High-Throughput In Vitro Metabolism Assay and LC−HRMS Metabolite Identification

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