Determination of cellular biodistribution of Cu, Ru and Pt in human tumor and non tumor breast cells by Inductively Coupled Plasma Mass Spectrometry

Abstract

The present project focuses in the determination of low concentrations of copper, ruthenium and platinum in breast human tumor and non-tumor cells lines using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The metal complexes containing clotrimazole (CTZ) and triphenylphosphine (PPh3)ligands were synthesized and present the following formulas: [Cu(CTZ)2(PPh3)2]+, [Ru(p-cym)(CTZ)(PPh3)Cl]+ and cis-[Pt(CTZ)(PPh3)2Cl]+. Initially, will be evaluated the cytotoxic activity of the complexes against breast tumor cell line (MDA-MB-231) and non-tumor (MCF-10A), also will be determined the cell death mechanism by cycle cell analysis and apoptosis assay. Posteriorly, the cells will be treated with the complexes for the determination of total metal concentration by ICP-MS, evaluating the following parameters: time, concentration, digestion of the sample assisted by microwave radiation, using different temperatures and HNO3 concentrations. These parameters will be evaluate to optimize and establish the most efficient treatment for the samples, that will be used for the determination of metal concentration in six isolates components of the cell: cytosol, plasma membrane, nucleus, cytoskeleton, DNA and finally mitochondria. The obtained results will be compared to each other and with reported data in the literature, to better understand the sample treatment and determine the cell distribution of the metal responsible for their biological properties in order to explore their mechanisms of action.