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The current study highlights the apoptotic activity of copper oxide (CuO) nanoparticles functionalized by Glutamic acid and conjugated by thiosemicarbazone (TSC) toward human breast cancer (MCF-7) and normal (HEK293) cell lines. To this aim, the co-precipitation method was used for preparation the CuO nanoparticles. Afterward, the CuO nanoparticles functionalized by glutamic acid. After that the functionalized copper oxide nanoparticles (CuO@Glu) conjugated to thiosemicarbazone. In next step, the final nanoparticle product (CuO@Glu/TSC) was characterized by physico-chemical techniques including Fourier transform infrared (FT-IR) spectroscopy, Scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Energy dispersive X-ray analysis (EDX), zeta potential analysis and dynamic light scattering (DLS). The effects of in vitro cell viability in CuO@Glu/TSC nanoparticles showed the anti-proliferative properties with a dose-dependent manner (IC50?=?133.97 µg/ml). The IC50 of CuO@Glu/TSC on normal cell line was 230.2 µg/ml. This IC50 deference shows high cytotoxicity of CuO@Glu/TSC nanoparticles on tumor cells and low cytotoxicity on non-tumorigenic cells (HEK293) and is considered as an important aspect for this nanoparticles. Also, CuO@Glu/TSC nanoparticles had efficient effects in inhibiting the growth of breast cancer cell line (MCF-7). In addition, the CuO@Glu/TSC nanoparticle induced apoptosis symptoms which were assessed by Caspase-3 activation assay, Annexin V/ propidium iodide flow cytometry, and Hoechst 33258 staining. Further, Bax and Bcl-2 genes expression was estimated by real time PCR. The expression of Bax increased 1.69 fold, while the expression of Bcl-2 decreased 0.6 fold. The results of the current study propose that CuO@Glu/TSC nanoparticles reveal effective anti-cancer activity against breast cancer cell line.

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