(1) identification of cell death mechanisms in the presence of Fe3O4 nanoparticles; 

(2) evaluating the influence of these nanoparticles in the formation and development of tumors.

(1) Morphological, phenotypic, genotypic and functional characterization in vitro of solid tumors in the presence of magnetite nanoparticles (Fe3O4); 

(2) Evaluation of the mechanisms of cell death induced by nanoparticles; 

(3) Identification in vitro of the characteristic markers resulting from the interaction of tumor cells exposed to nanoparticles with the tumor microenvironment; 

(4) The in vivo evaluation of the ability of tumor cells exposed to nanoparticles to form and develop tumors

During the process of tumorigenesis, tumor cells proliferate uncontrollably, evading the regulatory mechanisms of cell death. Several types of cell death have been described: apoptosis (type I), cell death associated with autophagy (type II), necrosis (type III), mitotic catastrophe, anchorage-dependent mechanisms – anoikis, excitotoxicity, Wallerian degeneration, skin cornification. This project proposes to investigate a possible new mechanism for inducing tumor cell death in vitro and in vivo – enucleation. The influence of Fe3O4 nanoparticles in colloidal suspensions on human tumor cell lines will be tested. Magnetic particles (MNPs) will be synthesized by combustion and coated with a double layer of oleic acid, interacting with tumor cells. In vitro studies will be focused on morphological and ultrastructural changes, functional studies, immunophenotypic markers and gene expression induced in tumor cells and resulting nuclei, using a modern methodology. Through in vivo models, the ability of tumor cells treated with magnetic nanoparticles to form tumors in immunodeficient mice will be detected, followed by the characterization of the generated tumors and associated metastases. This project will provide evidence regarding enucleation as a potential mechanism of tumor cell death, generating new directions in tumor biology research and the development of therapeutic agents capable of exploiting this behavior.

• The method of highlighting the effect of MNP on normal and tumor cells by SEM (Fig. 1)
• The method of highlighting the effect of MNP on normal and tumor cells by TEM (Fig. 2 and Fig. 3)
• The flow cytometric method for the analysis of changes induced by MNP at the cellular level (Fig. 4)
• The immunocytochemical method for morphological and immunophenotypic evidence of MNP-induced changes at the cellular level (Fig. 5)
• PBMC isolation protocols;
• Protocols for tumor cell-tumor microenvironment co-culture systems;
• Panel of characteristic markers of tumor cells treated with MNPs after interaction with immune cells;
• Protocols for GFP labeling of tumor cells;
• Protocols for tumor formation in immunosuppressed mice (Fig. 6);

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