New in vitro approaches to explore cellular and molecular events related to carcinogenesis

The integrated experimental approaches presented here have the potential to reveal the impairment of critical cellular and molecular events related to mutagenesis and carcinogenesis induced by exposure to specific chemical or physical agents and to appraise the chain of events leading to those effects. In particular, flow cytometry combined with FISH provides a better understanding of the mechanisms of DNAdamage induction. FISH with centromere DNA probes allowed the assessment of ratio of clastogenic vs aneugenic events. In addition, the FISH using multiple fluorescent chromosome probes was exploited to investigate the relatively propensity of each chromosome to loss or non-disjunction at mitosis in relation to size and DNA content. Furthermore, flow cytometry can also be applied in the study of molecular mechanisms associated with cell death. In this area flow cytometry offers unique opportunities and is even more advantageous compared to the alternative technique. By virtue of the possibility of correlating measurements of multiple parameters on the same cells, the methodology allows one to detect cells undergoing apoptosis with respect to their cell cycle position or to study the relationship between apoptosis and a particular function of the cell or cellular organelles.