BALB/c 3T3 cell transformation assay for the prediction of carcinogenic potential of chemicals and environmental mixtures

Mascolo, Maria Grazia; Perdichizzi, Stefania; Rotondo, Francesca; Morandi, Elena; Guerrini, Angela; Silingardi, Paola; Vaccari, Monica; Grilli, Sandro; Colacci, Annamaria

doi:10.1016/j.tiv.2010.03.003

Cited by 33 publications

(28 citation statements)

References 31 publications

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“…Because cell transformation is an event that is related with this carcinogenic capacity, we evaluated whether a metal mixture of 2 μ M NaAsO 2 , 2 μ M CdCl 2 , and 5 μ M Pb(C 2 H 3 O 2 ) 2 ·3H 2 O could transform Balb/c 3T3 cells. These cells have recently been mentioned as good model to the study of mixtures [37]. We evaluated the effects of the mixture as initiator, promoter, and both and determined the role of oxidative stress in transformation process.…”

Section: Discussionmentioning

confidence: 99%

Role of Oxidative Stress in Transformation Induced by Metal Mixture

Škandík

Rojas

Valverde

2011

Oxidative Medicine and Cellular Longevity

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Metals are ubiquitous pollutants present as mixtures. In particular, mixture of arsenic-cadmium-lead is among the leading toxic agents detected in the environment. These metals have carcinogenic and cell-transforming potential. In this study, we used a two step cell transformation model, to determine the role of oxidative stress in transformation induced by a mixture of arsenic-cadmium-lead. Oxidative damage and antioxidant response were determined. Metal mixture treatment induces the increase of damage markers and the antioxidant response. Loss of cell viability and increased transforming potential were observed during the promotion phase. This finding correlated significantly with generation of reactive oxygen species. Cotreatment with N-acetyl-cysteine induces effect on the transforming capacity; while a diminution was found in initiation, in promotion phase a total block of the transforming capacity was observed. Our results suggest that oxidative stress generated by metal mixture plays an important role only in promotion phase promoting transforming capacity.

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Section: Discussionmentioning

confidence: 99%

Role of Oxidative Stress in Transformation Induced by Metal Mixture

Škandík

Rojas

Valverde

2011

Oxidative Medicine and Cellular Longevity

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“…The filtered insoluble solid was dissolved in tetrahydrofuran, and then dried by MgSO 4 . The product, 3,6-di(4-vinylpyridine) carbazole, was collected by recrystallization from tetrahydrofuran filtrate [28]. In the preparation of BMVC probes, BMVC stock solution was dissolved in dimethyl sulfoxide (DMSO) at 2 mg/ml, which was further diluted to a working concentration of 2 μM when preparing the BMVC probes.…”

Section: Methodsmentioning

confidence: 99%

“…To investigate the capacity of BMVC for detecting cell transformation during carcinogenesis, a system of inducible cell transformation is important for in vitro analysis [28]–[30]. A BALB/c 3T3 cell transformation assay was performed according to a modified protocol as previously described [22]–[26].…”

Section: Methodsmentioning

confidence: 99%

Detection of Cell Carcinogenic Transformation by a Quadruplex DNA Binding Fluorescent Probe

et al. 2014

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Cancer can be easily treated when found early. A probe capable of detecting cell transformation may increase the success rate of early diagnosis of cancer. In this report we have tested the ability of a fluorescent, quadruplex DNA binding probe, 3,6-bis(1-methyl-4- vinylpyridinium) carbazole diiodide (BMVC), to detect cell transformation in vitro. BMVC was applied to living cells in several different models of cell transformation, and the fluorescence signals of BMVC were measured. The degrees of cell transformation in these models were characterized by alterations in cellular morphological phenotype and subcellular organization. When BMVC probes were applied, the number of BMVC-positive cells increased in accordance with the degree of transformation. BMVC was capable of significantly detecting formation of foci, increased cellular motility, cell proliferation, cell apoptosis, anchorage-independent growth, and increased invasiveness of transformed cells. These results demonstrate the ability of BMVC probes to detect cell transformation and indicate that BMVC is of promise for use as a probe in early cancer detection.

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“…The transformation assay was performed as previously reported [13,14] with minor modifications. Cells were seeded at a density of 3 × 10 4 cells/60 mm dish, incubated for 48 h and then exposed to the PM 2.5 extracts at concentrations ranging from 1.5 to 12 m 3 equivalents.…”

Section: Morphological Transformation Assaymentioning

confidence: 99%

Alternative Testing Methods for Predicting Health Risk from Environmental Exposures

et al. 2014

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Alternative methods to animal testing are considered as promising tools to support the prediction of toxicological risks from environmental exposure. Among the alternative testing methods, the cell transformation assay (CTA) appears to be one of the most appropriate approaches to predict the carcinogenic properties of single chemicals, complex mixtures and environmental pollutants. The BALB/c 3T3 CTA shows a good degree of concordance with the in vivo rodent carcinogenesis tests. Whole-genome transcriptomic profiling is performed to identify genes that are transcriptionally regulated by different kinds of exposures. Its use in cell models representative of target organs may OPEN ACCESS Sustainability 2014, 6 5266 help in understanding the mode of action and predicting the risk for human health. Aiming at associating the environmental exposure to health-adverse outcomes, we used an integrated approach including the 3T3 CTA and transcriptomics on target cells, in order to evaluate the effects of airborne particulate matter (PM) on toxicological complex endpoints. Organic extracts obtained from PM 2.5 and PM 1 samples were evaluated in the 3T3 CTA in order to identify effects possibly associated with different aerodynamic diameters or airborne chemical components. The effects of the PM 2.5 extracts on human health were assessed by using whole-genome 44 K oligo-microarray slides. Statistical analysis by GeneSpring GX identified genes whose expression was modulated in response to the cell treatment. Then, modulated genes were associated with pathways, biological processes and diseases through an extensive biological analysis. Data derived from in vitro methods and omics techniques could be valuable for monitoring the exposure to toxicants, understanding the modes of action via exposure-associated gene expression patterns and to highlight the role of genes in key events related to adversity.

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BALB/c 3T3 cell transformation assay for the prediction of carcinogenic potential of chemicals and environmental mixtures

Cited by 33 publications

References 31 publications

Role of Oxidative Stress in Transformation Induced by Metal Mixture

Role of Oxidative Stress in Transformation Induced by Metal Mixture

Detection of Cell Carcinogenic Transformation by a Quadruplex DNA Binding Fluorescent Probe

Alternative Testing Methods for Predicting Health Risk from Environmental Exposures

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