Ricerc@Sapienza

Lindane (or gamma-HCH) is a synthetical organochlorine pesticide, widely used in the past but further banned due to its great toxicity and environmental persistence. Industrial waste sites and illegal dumping further contributed to the environment contamination and bioaccumulation, thus leading to an increased susceptibility of exposed people to various degenerative diseases. This suggest the need of preventive and care interventions to maintain human health. Among them, chemoprevention is aimed at counteracting chemical-induced DNA damage thus blocking cancer development and progression.
In line with this evidence, in this project lindane genotoxicity will be characterized in order to both clarify its toxicity mechanisms and identify suitable genoprotective natural substances for further chemopreventive applications. The H2AX histone assay, a novel method for genotoxicity detection, which measures the H2AX phosphorilation (namely gamma-H2AX) as a results of DNA damage and instability, has been selected for this study, being affordable and versatile along with sensitive and specific compared the standard methodologies. Two in vitro cell models, i.e. the human hepatoma HepG2 cell line and primary human lymphocytes will be used. Validating the assay in primary lymphocytes can also support a further application of this assay in biomonitoring studies. Lindane-induced gamma-H2AX will be detected through flow cytometric measurements, western blotting and immunofluorescence analysis, thus allowing to rapidly analyze large numbers of samples and to further quantify and localize the protein. Moreover, combining gamma-H2AX with the phosphorylated histone H3 allows to characterize aneugenic or clastogenic genotoxicity mechanisms. Feasibility of this project is guaranteed by the skills of Dr. Gullì acquired during his internship for thesis degree and further improved during the PhD in Toxicology and by the availability of adequate laboratory infrastructures and equipments.