An international validation study of a Bhas 42 cell transformation assay for the prediction of chemical carcinogenicity

Sakai, Ayako; Sasaki, Kiyoshi; Hayashi, Kumiko; Muramatsu, Dai; Arai, Sadahiko; Endou, Nobuko; Kuroda, Sachiko; Poth, Albrecht; Bohnenberger, Susanne; Kunkelmann, Thorsten; Asakura, Masanori; Hirose, Hideki; Ishii, Nana; Mizuhashi, Fukutaro; Kasamoto, Sawako; Nagai, Miho; Pant, Kamala; Bruce, Shannon; Sly, Jamie; Yamazaki, Shojiro; Umeda, Makoto; Tanaka, Noriho

doi:10.1016/j.mrgentox.2011.07.006

Cited by 36 publications

(32 citation statements)

References 13 publications

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“…In this study, the numbers of transformed foci obtained in control culture and in treated culture (MCA in the initiation assay and TPA in the promotion assay) were in agreement with those previously published [21,22]. At the beginning of our study, the Bhas 42 CTA had not yet been tested with particle-type substances.…”

Section: Discussionsupporting

confidence: 91%

In vitro cell transformation induced by synthetic amorphous silica nanoparticles

Fontana

Kirsch

Seidel

et al. 2017

Mutation Research/Genetic Toxicology and Environmental Mutagene

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International audienceSynthetic amorphous silica nanoparticles (SAS) are among the most widely produced and used nanomaterials, but little is known about their carcinogenic potential. This study aims to evaluate the ability of four different SAS, two precipitated, NM-200 and NM-201, and two pyrogenic, NM-202 and NM-203, to induce the transformation process. For this, we used the recently developed in vitro Bhas 42 cell transformation assay (CTA). The genome of the transgenic Bhas 42 cells contains several copies of the v-Ha-ras gene, making them particularly sensitive to tumor-promoter agents. The Bhas 42 CTA, which includes an initiation assay and a promotion assay, was validated in our laboratory using known soluble carcinogenic substances. Its suitability for particle-type substances was verified by using quartz Min-U-Sil 5 (Min-U-Sil) and diatomaceous earth (DE) microparticles. As expected given their known transforming properties, Min-U-Sil responded positively in the Bhas 42 CTA and DE responded negatively. Transformation assays were performed with SAS at concentrations ranging from 2 µg/cm 2 to 80 µg/cm 2. Results showed that all SAS have the capacity to induce transformed foci, interestingly only in the promotion assay, suggesting a mode of action similar to tumor-promoter substances. NM-203 exhibited transforming activity at a lower concentration than the other SAS. In conclusion, this study showed for the first time the transforming potential of different SAS, which act as tumor-promoter substances in the Bhas 42 model of cell transformation.Les nanoparticules de silice amorphes de synthèse (SAS) sont les nanomatériaux les plus produits et les plus utilisés dans le monde, mais leur potentiel cancérogène est peu étudié. Cette étude avait pour but d’évaluer la capacité de quatre différentes SAS, deux sous forme précipitée, NM-200 et NM-201 et deux sous forme pyrogénée, NM-202 et NM-203, d’induire des processus de cancérogénèse. Pour cela, nous avons utilisé l’essai de transformation sur les cellules Bhas 42 (Bhas 42 CTA) qui a été récemment développé. Le génome des cellules Bhas 42 transgéniques contient plusieurs copies du gène v-Ha-ras, ce qui leur confère les caractéristiques de cellules « initiées » et les rend particulièrement sensibles aux agents promoteurs de tumeur. Le Bhas 42 CTA, qui inclut un essai d’initiation et un essai de promotion, a été validé dans notre laboratoire en utilisant des agents cancérogènes solubles connus. Son adaptabilité pour des substances de type particule a été vérifié en utilisant des microparticules de quartz Min-U-Sil 5 (Min-U-Sil) et de terre de diatomée (DE). Comme attendu d’après leur propriété transformante connue, le Min-U-Sil a répondu positivement et la DE négativement au Bhas 42 CTA. Les essais de transformation ont été réalisés à des concentrations de SAS s’étalant de 2 µg/cm2 à 80 µg/cm2. Les résultats montrent que toutes les SAS ont la capacité d’induire des foyers de transformation et de manière intéressante, uniquement dans l’essai de promotion, suggérant...

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Section: Discussionsupporting

confidence: 91%

In vitro cell transformation induced by synthetic amorphous silica nanoparticles

Fontana

Kirsch

Seidel

et al. 2017

Mutation Research/Genetic Toxicology and Environmental Mutagene

View full text Add to dashboard Cite

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“… Schematic representation of initiation and promotion assays in the Bhas 42 cell transformation [Sakai et al, ]. MEM + 10% FBS was used for the first culture after thawing for population expansion.…”

Section: Methodsmentioning

confidence: 99%

“…These assays were performed at the concentration range of 0-40 lg mL 21 for both CSC-D and CSC-W. CSC-W did not elevate the average number of transformed foci at any of the tested concentrations when compared to the corresponding control (i.e., deionized water only). In contrast, significant increases in the number of transformed foci was observed when Bhas 42 [Sakai et al, 2011]. MEM 1 10% FBS was used for the first culture after thawing for population expansion.…”

Section: Initiation Activity Of Csc-w and Csc-d In Bhas 42 Cellsmentioning

confidence: 97%

Bhas 42 cell transformation activity of cigarette smoke condensate is modulated by selenium and arsenic

Han

Pant

Bruce

et al. 2016

Environ and Mol Mutagen

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Cigarette smoking remains a major health risk worldwide. Development of newer tobacco products requires the use of quantitative toxicological assays. Recently, v-Ha-ras transfected BALB/c3T3 (Bhas 42) cell transformation assay was established that simulates the two-stage animal tumorigenesis model and measures tumor initiating and promoting activities of chemicals. The present study was performed to assess the feasibility of using this Bhas 42 cell transformation assay to determine the initiation and promotion activities of cigarette smoke condensate (CSC) and its water soluble fraction. Further, the modulating effects of selenium and arsenic on cigarette smoke-induced cell transformation were investigated. Dimethyl sulfoxide (DMSO) and water extracts of CSC (CSC-D and CSC-W, respectively) were tested at concentrations of 2.5-40 µg mL(-1) in the initiation or promotion assay formats. Initiation protocol of the Bhas 42 assay showed a 3.5-fold increase in transformed foci at 40 µg mL(-1) of CSC-D but not CSC-W. The promotion phase of the assay yielded a robust dose response with CSC-D (2.5-40 µg mL(-1)) and CSC-W (20-40 µg mL(-1)). Preincubation of cells with selenium (100 nM) significantly reduced CSC-induced increase in cell transformation in initiation assay. Co-treatment of cells with a sub-toxic dose of arsenic significantly enhanced cell transformation activity of CSC-D in promotion assay. The results suggest a presence of both water soluble and insoluble tumor promoters in CSC, a role of oxidative stress in CSC-induced cell transformation, and usefulness of Bhas 42 cell transformation assay in comparing tobacco product toxicities and in studying the mechanisms of tobacco carcinogenesis.

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“…The results of the inter-laboratory studies have recently been published, and the data demonstrate that the Bhas 42 assay is reproducible between laboratories (22). …”

Section: Recent Validation Workmentioning

confidence: 99%

Cell transformation assays for prediction of carcinogenic potential: state of the science and future research needs

Creton¹,

Aardema²,

Carmichael

et al. 2011

Mutagenesis

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Cell transformation assays (CTAs) have long been proposed as in vitro methods for the identification of potential chemical carcinogens. Despite showing good correlation with rodent bioassay data, concerns over the subjective nature of using morphological criteria for identifying transformed cells and a lack of understanding of the mechanistic basis of the assays has limited their acceptance for regulatory purposes. However, recent drivers to find alternative carcinogenicity assessment methodologies, such as the Seventh Amendment to the EU Cosmetics Directive, have fuelled renewed interest in CTAs. Research is currently ongoing to improve the objectivity of the assays, reveal the underlying molecular changes leading to transformation and explore the use of novel cell types. The UK NC3Rs held an international workshop in November 2010 to review the current state of the art in this field and provide directions for future research. This paper outlines the key points highlighted at this meeting.

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An international validation study of a Bhas 42 cell transformation assay for the prediction of chemical carcinogenicity

Cited by 36 publications

References 13 publications

In vitro cell transformation induced by synthetic amorphous silica nanoparticles

In vitro cell transformation induced by synthetic amorphous silica nanoparticles

Bhas 42 cell transformation activity of cigarette smoke condensate is modulated by selenium and arsenic

Cell transformation assays for prediction of carcinogenic potential: state of the science and future research needs

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