The transformics assay: first steps for the development of an integrated approach to investigate the malignant cell transformation in vitro

Mascolo, Maria Grazia; Perdichizzi, Stefania; Vaccari, Monica; Rotondo, Francesca; Zanzi, Cristina; Grilli, Sandro; Paparella, Martin; Jacobs, Miriam N.; Colacci, Annamaria

doi:10.1093/carcin/bgy037

Cited by 32 publications

(49 citation statements)

References 49 publications

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“…Thus, in order to improve the use of this assay in an IATA for carcinogenesis, we developed a method combining the in vitro CTA and next‐generation sequencing‐based DNA methylation profiling analysis (MBD‐Seq) to elucidate the molecular steps leading to in vitro malignant transformation. In an approach similar to that in our study, Mascolo et al [] developed the transformics method, which combines the in vitro CTA and transcriptomics. In this study, the in vitro Bhas 42 CTA was performed with three nongenotoxic carcinogens (cadmium chloride, methyl carbamate, and lithocholic acid) whose genotoxicity and carcinogenicity have been confirmed in a wide range of test systems.…”

Section: Discussionmentioning

confidence: 99%

Genome‐wide DNA methylation changes in transformed foci induced by nongenotoxic carcinogens

Hwang

Yeom

Eom

et al. 2019

Environ and Mol Mutagen

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In vitro cell transformation assays (CTA) have been proposed as a method to identify possible nongenotoxic carcinogens. However, the current protocols do not provide information on the mechanism of action of the test articles. In this study, we combined an in vitro Bhas 42 CTA and sequencing‐based DNA methylation profiling analysis to elucidate the carcinogenic mechanism associated with nongenotoxic carcinogens. Three nongenotoxic carcinogens were evaluated: cadmium chloride, methyl carbamate, and lithocholic acid. Methylation profiles were generated for the two nongenotoxic carcinogens (cadmium chloride and lithocholic acid) that were positive in Bhas 42 CTA. Methyl carbamate did not exhibit any promoter activity. Approximately 9.8% of all differentially methylated regions (DMRs) identified in cadmium chloride‐induced transformed foci overlapped with DMRs in lithocholic acid‐induced transformed foci. Interestingly, overlapping DMRs showed more hypermethylation than individual DMRs. In addition, the DMRs in CpG island elements common to both nongenotoxic carcinogens showed considerably more bias toward hypermethylated DMRs than those unique to either cadmium chloride or lithocholic acid. Pathway enrichment analysis revealed that genes harboring hypermethylated DMRs were significantly enriched in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways including pathways in cancer, basal cell carcinoma, and Wnt signaling. The genes harboring hypomethylated DMRs were significantly related to mRNA surveillance pathway, RNA transport, and autophagy. Taken together, our preliminary results on genome‐wide methylation analysis of cell clones from nongenotoxic carcinogen‐induced foci could be exploited for CTAs improvement, but further research will be required to standardize and assess the specificity and sensitivity of this combined approach. Environ. Mol. Mutagen. 2019. © 2019 Wiley Periodicals, Inc.

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Section: Discussionmentioning

confidence: 99%

Genome‐wide DNA methylation changes in transformed foci induced by nongenotoxic carcinogens

Hwang

Yeom

Eom

et al. 2019

Environ and Mol Mutagen

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“…On the basis of our results, we speculated that the binding of PM components, such as PAHs, to the Aryl hydrocarbon Receptor (AhR) triggers an immune-mediated response, leading to the activation of molecular pathways sustaining inflammation, through the release of several cytokines as signal mediators (Mascolo et al, 2018). Cytokines play a key role in the innate immune response against pathogens or toxic substances and have a complex regulatory effect on the inflammation and immune system.…”

Section: Introductionmentioning

confidence: 93%

“…The toxicological properties increase linearly with increase in the PAHs concentration. Low concentrations trigger detoxification pathways, whilst higher concentrations activate an aberrant immune response, leading to genomic instability (Colacci et al, 2014;Vaccari et al, 2015;Mascolo et al, 2018;Serra et al, 2019). The entire process is orchestrated by the AhR.…”

Section: Pm and The Receptors Interplaying In Adverse Outcomes By Chementioning

confidence: 99%

“…It is the molecular target for dioxins, dioxin-like chemicals as well as PAHs, mediating the metabolic activation of xenobiotics and the response to environmental exposures. The complex AhR-ARNT binds specific DNA sequences, triggering the AhR signaling pathway, which leads to the transcription of the CYP1A1 and CYP1B1 genes, whose proteins play the main role in the bioactivation of several xenobiotics (Mascolo et al, 2018). The AhR is the only PAS protein that is activated by the ligand.…”

Section: Pm and The Receptors Interplaying In Adverse Outcomes By Chementioning

confidence: 99%

“…We have also found that the toxicological behavior of PM was strictly related to the content of PM components, such as the polycyclic aromatic hydrocarbons (PAHs). Indeed, PM can induce cell malignant transformation in vitro only when the concentration of PAHs exceeds the cells ability to implement its adaptive or detoxicant response (Vaccari et al, 2015;Mascolo et al, 2018;Serra et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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The transformics assay: first steps for the development of an integrated approach to investigate the malignant cell transformation in vitro

Abstract: The transformics method was developed to support the integrated testing strategy for carcinogenesis. Results showed the main role of the immune system in the transformation by 3-MCA, with initiating events related to non-genotoxic mechanisms, suggesting the involvement of the AhR receptor.

Cited by 32 publications

References 49 publications

Genome‐wide DNA methylation changes in transformed foci induced by nongenotoxic carcinogens

Genome‐wide DNA methylation changes in transformed foci induced by nongenotoxic carcinogens

The Secretive Liaison of Particulate Matter and SARS-CoV-2. A Hypothesis and Theory Investigation

Molecular insights into acetyl triethyl citrate (ATEC) induced toxic effect in HepG2 cells based on multi‐omics integrative analysis

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