Nvestigation of the synthesis, characterization and usability of TNF-doxorubicin loaded gold nanoparticles in breast cancer treatment under in vitro conditions

In this study, a nanotechnological drug carrier was developed, which could be utilized as an innovative approach in the treatment of breast cancer. Gold nanoparticles (GNP) were modified with tumor necrosis factor-alpha (TNF) to specifically target tumor cells, and then loaded with doxorubicin (Dox) to create a therapeutic molecule. The anti-cancer effect of this newly developed drug carrier nanoparticle was tested on MCF-7 and AMJ-13 breast cancer cell lines, as well as the healthy cell line REF. Firstly, the characterization of the created drug carrier nanoparticle was conducted using ultraviolet-visible spectroscopy (UV-Vis spectroscopy), transmission electron microscopy (TEM), and ELISA techniques. Subsequently, the cytotoxic effect of TNFα, GNP, Dox, GNP-TNFα, and GNP-TNFα-Dox test compounds on breast cancer cells was evaluated using MTT and clonogenicity tests. The IC50 doses of each test compound were determined. The potential of inducing apoptosis in MCF-7, AMJ-13, and REF cells by the test compounds was investigated by observing nuclear morphology under a fluorescence microscope using the AO and EtBr double staining technique. Additionally, the potential of inducing autophagy in breast cancer cells by each test compound was measured by flow cytometry using the LC3 protein level. As a result, it was found that the GNP-TNFα-Dox complex from the prepared nanoparticles exhibited a more effective cytotoxicity and anticancer effect, particularly compared to the Dox treatment currently used in clinics. Furthermore, it was observed that GNP-TNFα-Dox complexes were significantly cytotoxic to breast cancer cell lines in comparison to the control group, accomplishing this through both apoptosis and autophagy induction. These results demonstrated that GNP-TNFα-Dox complexes could constitute an effective anticancer treatment for the targeted cancer therapy approach, which is one of the main objectives of the thesis