Abstract
The identification of cancer preventive or therapeutic substances as well as carcinogenic risk assessment of chemicals is nowadays mostly dependent on animal studies. In vitro cell transformation assays mimic different stages of the in vivo neoplastic process and represent an excellent alternative to study carcinogenesis and therapeutic options. In the BALB/c-3T3 two-stage transformation assay cells are chemically transformed by treatment with MCA and TPA, along with the final Giemsa staining of morphological aberrant foci. In addition to the standard method we can show, that it is possible to apply other chemicals in parallel to identify potential preventive or therapeutic substances during the transformation process. Furthermore, we successfully combined the BALB/c cell transformation assay with several endpoint applications for protein analysis (immunoblot, subcellular fractionation and immunofluorescence) or energy parameter measurements (glucose and oxygen consumption) to elucidate cancer mechanisms in more detail. In our opinion the BALB/c cell transformation assay proves to be an excellent model to investigate alterations in key proteins or energy parameters during the different stages of transformation as well as therapeutic substances and their mode of action.
Highlights
Several modifications of the classical method had been carried out, like the use of medium with pH 6.714,15 or an initiation-promotion protocol[16]
Despite the identification of potential tumor initiators and promotors by using cell transformation assays as standard toxicological methods we further improved the BALB-Cell transformation protocol (CTA) for mechanistic cancer research
Reproducibility and efficiency of the BALB-CTA protocol was shown by the effects of well-known carcinogens
Summary
Several modifications of the classical method had been carried out, like the use of medium with pH 6.714,15 or an initiation-promotion protocol[16]. Different improvements of the standard protocol were proposed, like a two-stage assay with treatment of suspected carcinogens followed by a known tumor promotor[20], the use of the new developed Bhas 42 cell line (BALB/c-3T3 transfected with v-Ha-ras)[21,22,23] or the combination of the BALB-CTA with microarray-based toxicogenomics[24]. Despite the identification of potential tumor initiators and promotors by using cell transformation assays as standard toxicological methods we further improved the BALB-CTA for mechanistic cancer research.
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