A Bhas 42 cell transformation assay on 98 chemicals: The characteristics and performance for the prediction of chemical carcinogenicity

Sakai, Ayako; Sasaki, Kiyoshi; Muramatsu, Dai; Arai, Sadahiko; Endou, Nobuko; Kuroda, Sachiko; Hayashi, Kumiko; Lim, Yeon‐Mi; Yamazaki, Shojiro; Umeda, Makoto; Tanaka, Noriho

doi:10.1016/j.mrgentox.2010.07.007

Cited by 46 publications

(45 citation statements)

References 17 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%

“…A validation study covering 98 chemicals established an 84% specificity of the Bhas 42 CTA towards carcinogens and non-carcinogens [21].…”

Section: Discussionmentioning

confidence: 99%

“…DE treatment did not induce cell transformation, but decreased the number of transformed foci in the initiation and promotion assays. Other non-carcinogen chemicals such as L-ascorbic acid or caffeine have been found to lower the number of spontaneous transformed Bhas 42 foci [21]..…”

Section: Discussionmentioning

confidence: 99%

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In vitro cell transformation induced by synthetic amorphous silica nanoparticles

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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%

“…A validation study covering 98 chemicals established an 84% specificity of the Bhas 42 CTA towards carcinogens and non-carcinogens [21].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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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“…Recently, we applied the Bhas 42 cell transformation assay to 98 chemicals including 52 known carcinogens and 37 noncarcinogens to evaluate its performance for the detection of chemical carcinogenicity [60]. The indices of the performance of in vitro data to predict rodent carcinogenicity are shown in Table 2 78%, sensitivity 73%, specificity 84%, positive predictivity 86%, negative predictivity 69%, false negative 16%, and false positive 27%.…”

Section: Assessment Of the Bhas 42 Cell Transformation Assay For Carcmentioning

confidence: 99%

Transformation Assay in Bhas 42 Cells: A Model Using Initiated Cells to Study Mechanisms of Carcinogenesis and Predict Carcinogenic Potential of Chemicals

Sasaki

Umeda

Sakai

et al. 2015

Journal of Environmental Science and Health, Part C

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Transformation assays using cultured cells have been applied to the study of carcinogenesis. Although various cell systems exist, few cell types such as BALB/c 3T3 subclones and Syrian hamster embryo cells have been used to study chemically induced two-stage carcinogenesis. Bhas 42 cells were established as a clone by the transfection with the v-Ha-ras gene into mouse BALB/c 3T3 A31-1-1 cells and their subsequent selection based on their sensitivity to 12-O-tetradecanoylphorbol-13-acetate. Using Bhas 42 cells, transformed foci were induced by the treatment with nongenotoxic carcinogens, most of which act as tumor promoters. Therefore, Bhas 42 cells were considered to be a model of initiated cells. Subsequently, not only nongenotoxic carcinogens but also genotoxic carcinogens, most of which act as tumor initiators, were found to induce transformed foci by the modification of the protocol. Furthermore, transformation of Bhas 42 cells was induced by the transfection with genes of oncogenic potential. We interpret this high sensitivity of Bhas 42 cells to various types of carcinogenic stimuli to be related to the multistage model of carcinogenesis, as the transfection of v-Ha-ras gene further advances the parental BALB/c 3T3 A31-1-1 cells toward higher transforming potential. Thus, we propose that Bhas 42 cells are a novel and sensitive cell line for the analysis of carcinogenesis and can be used for the detection of not only carcinogenic substances but also gene alterations related to oncogenesis. This review will address characteristics of Bhas 42 cells, the transformation assay protocol, validation studies, and the various chemicals tested in this assay.

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“…1 ) [ 11 ]. The ability of the Bhas 42 assay to detect genotoxic and non-genotoxic carcinogens was evaluated on 98 chemicals [ 10 ]. The assay was compared with the BALB/c 3T3 transformation assay [ 12 , 13 ], and pre-validated and validated in interlaboratory studies using two protocols, for the 6-and 96-well plates [ 14 -17 ].…”

Section: Introductionmentioning

confidence: 99%

Bhas 42 Cell Transformation Assay for Genotoxic and Non-Genotoxic Carcinogens

Sasaki

Huk

Yamani

et al. 2014

Genotoxicity and DNA Repair

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Cell transformation assays (CTAs) are in vitro carcinogenicity tests measuring morphological transformation of cells either as transformed colonies (SHE cells) or foci (C3H/10T1/2 and BALB/c 3T3 including Bhas 42 cells) derived from a single cell. CTAs such as Bhas 42 CTA can detect both genotoxic and nongenotoxic carcinogens. When used as an initiation assay to test tumor-initiating activity, cells at low density are treated with a test chemical for 3 days, whereas a promotion assay to test for tumor-promoting activity, near-confl uent cells are treated with a test chemical for a period of 10 days. The Bhas 42 CTA has advantages compared with BALB/c 3T3 and other CTAs due to its simplicity, higher sensitivity, less time needed for assay performance, and robustness (exemplifi ed by its adaptation to a high-throughput method). The Bhas 42 CTA has been validated together with other CTAs and recommended for development of an OECD guideline. The assay has already been applied in testing various chemical and physical agents including particles and nanomaterials. Protocols for both 6 and 96-well plate formats of initiation and promotion assays are described in detail.

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A Bhas 42 cell transformation assay on 98 chemicals: The characteristics and performance for the prediction of chemical carcinogenicity

Cited by 46 publications

References 17 publications

In vitro cell transformation induced by synthetic amorphous silica nanoparticles

In vitro cell transformation induced by synthetic amorphous silica nanoparticles

Transformation Assay in Bhas 42 Cells: A Model Using Initiated Cells to Study Mechanisms of Carcinogenesis and Predict Carcinogenic Potential of Chemicals

Bhas 42 Cell Transformation Assay for Genotoxic and Non-Genotoxic Carcinogens

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